Self-replicating RNA molecule from hepatitis C virus

ABSTRACT

A unique HCV RNA molecule is provided having an enhanced efficiency of establishing cell culture replication. Novel adaptive mutations have been identified within the HCV non-structural region that improves the efficiency of establishing persistently replicating HCV RNA in cell culture. This self-replicating polynucleotide molecule contains, contrary to all previous reports, a 5′-NTR that can be either an A as an alternative to the G already disclosed and therefore provides an alternative to existing systems comprising a self-replicating HCV RNA molecule. The G-&gt;A mutation gives rise to HCV RNA molecules that, in conjunction with mutations in the HCV non-structural region, such as the G(2042)C/R mutations, possess greater efficiency of transduction and/or replication. These RNA molecules when transfected in a cell line are useful for evaluating potential inhibitors of HCV replication.

RELATED APPLICATIONS

[0001] This application is a divisional of U.S. Application Serial No.10/029,907, filed Dec. 21, 2001, which claims, as does the presentapplication priority to U.S. Provisional Application Serial No.60/257,857 filed on Dec. 22, 2000, the disclosures of all of which areincorporated by reference in their entirety.

FIELD OF THE INVENTION

[0002] The present invention relates generally to a HCV RNA moleculethat self-replicates in appropriate cell lines, particularly to aself-replicating HCV RNA construct having an enhanced efficiency ofestablishing cell culture replication.

BACKGROUND OF THE INVENTION

[0003] Hepatitis C virus (HCV) is the major etiological agent ofpost-transfusion and community-acquired non-A non-B hepatitis worldwide.It is estimated that over 200 million people worldwide are infected bythe virus. A high percentage of carriers become chronically infected andmany progress to chronic liver disease, so called chronic hepatitis C.This group is in turn at high risk for serious liver disease such asliver cirrhosis, hepatocellular carcinoma and terminal liver diseaseleading to death. The mechanism by which HCV establishes viralpersistence and causes a high rate of chronic liver disease has not beenthoroughly elucidated. It is not known how HCV interacts with and evadesthe host immune system. In addition, the roles of cellular and humoralimmune responses in protection against HCV infection and disease haveyet to be established.

[0004] Various clinical studies have been conducted with the goal ofidentifying pharmaceutical compounds capable of effectively treating HCVinfection in patients afflicted with chronic hepatitis C. These studieshave involved the use of interferon-alpha, alone and in combination withother antiviral agents such as ribavirin. Such studies have shown that asubstantial number of the participants do not respond to thesetherapies, and of those that do respond favorably, a large proportionwere found to relapse after termination of treatment. To date there areno broadly effective antiviral compounds for treatment of HCV infection.

[0005] HCV is an enveloped positive strand RNA virus in the Flaviviridaefamily. The single strand HCV RNA genome is of positive polarity andcomprises one open reading frame (ORF) of approximately 9600 nucleotidesin length, which encodes a linear polyprotein of approx. 3010 aminoacids. In infected cells, this polyprotein is cleaved at multiple sitesby cellular and viral proteases to produce structural and non-structural(NS) proteins. The structural proteins (C, E1, E2 and E2-p7) comprisepolypeptides that constitute the virus particle (Hijikata, M. et al.,1991, Proc. Natl. Acad. Sci. USA. 88, 5547-5551; Grakoui et al.,1993(a), J. Virol. 67, 1385-1395). The non-structural proteins (NS2,NS3, NS4A, NS4B, NS5A, NS5B) encode for enzymes or accessory factorsthat catalyze and regulate the replication of the HCV RNA genome.Processing of the structural proteins is catalyzed by host cellproteases (Hijikata, M. et al., 1991, Proc. Natl. Acad. Sci. USA. 88,5547-5551). The generation of the mature non-structural proteins iscatalyzed by two virally encoded proteases. The first is the NS2/3zinc-dependent metalloprotease which auto-catalyses the release of theNS3 protein from the polyprotein. The released NS3 contains a N-terminalserine protease domain (Grakoui et al., 1993(b), Proc Natl Acad Sci USA,90,10583-7; Hijikata, M. et al., 1993, J. Virol. 67, 4665-4675) andcatalyzes the remaining cleavages from the polyprotein. The releasedNS4A protein has at least two roles. First, forming a stable complexwith NS3 protein and assisting in the membrane localization of theNS3/NS4A complex (Kim et al., Arch Virol. 1999, 144, 329-343) andsecond, acting as a cofactor for NS3 protease activity. Thismembrane-associated complex, in turn catalyzes the cleavage of theremaining sites on the polyprotein, thus effecting the release of NS4B,NS5A and NS5B (Bartenschlager, R. et al., 1993, J. Virol., 67,3835-3844; Grakoui et al., 1993(a), J. Virol. 67,1385-1395; Hijikata, M.et al., 1993, J. Virol. 67, 4665-4675; Love, R. A. et al., 1996, Cell,87, 331-342; reviewed in Kwong et al., 1998 Antiviral Res., 40, 1-18).The C-terminal segment of the NS3 protein also harbors nucleosidetriphosphatase and RNA helicase activity (Kim et al., 1995, Biochem.Biophys. Res. Comm., 215, 160-166.). The function of the protein NS4B isunknown. NS5A, a highly phosphorylated protein, seems to be responsiblefor the Interferon resistance of various HCV genotypes (Gale Jr. et al.1997 Virology 230, 217; Reed et al., 1997 J. Virol. 71, 7187. NS5B is anRNA-dependent RNA polymerase (RdRp) that is involved in the replicationof HCV.

[0006] The open reading frame of the HCV RNA genome is flanked on its 5′end by a non-translated region (NTR) of approx. 340 nucleotides thatfunctions as the internal ribosome entry site (IRES), and on its 3′ endby a NTR of approximately 230 nucleotides. Both the 5′ and 3′ NTRs areimportant for RNA genome replication. The genomic sequence variance isnot evenly distributed over the genome and the 5′NTR and parts of the3′NTR are the most highly conserved portions. The authentic, highlyconserved 3′NTR is the object of U.S. Pat. No. 5,874,565 granted to Riceet al.

[0007] The cloned and characterized partial and complete sequences ofthe HCV genome have also been analyzed with regard to appropriatetargets for a prospective antiviral therapy. Four viral enzymeactivities provide possible targets such as (1) the NS2/3 protease; (2)the NS3/4A protease complex, (3) the NS3 Helicase and (4) the NS5BRNA-dependent RNA polymerase. The NS3/4A protease complex and the NS3helicase have already been crystallized and their three-dimensionalstructure determined (Kim et al., 1996, Cell, 87, 343; Yem et al.Protein Science, 7, 837,1998; Love, R. A. et al., 1996, Cell, 87,331-342; Kim et al., 1998, Structure, 6, 89; Yao et al., 1997 NatureStructural Biology, 4,463; Cho et al., 1998, J. Biol. Chem., 273,15045). The NS5B RNA dependent RNA polymerase has also been crystallizedto reveal a structure reminiscent of other nucleic acid polymerases(Bressanelli et al. 1999, Proc. Natl. Acad. Sci, USA 96, 13034-13039;Ago et al. 1999, Structure 7, 1417-1426; Lesburg et al. 1999, Nat.Struct. Biol. 6, 937-943).

[0008] Even though important targets for the development of a therapyfor chronic HCV infection have been defined with these enzymes and eventhough a worldwide intensive search for suitable inhibitors is ongoingwith the aid of rational drug design and HTS, the development of therapyhas one major deficiency, namely the lack of cell culture systems orsimple animal models, which allow direct and reliable propagation of HCVviruses. The lack of an efficient cell culture system is still the mainreason to date that an understanding of HCV replication remains elusive.

[0009] Although flavi- and pestivirus self-replicating RNAs have beendescribed and used for the replication in different cell lines with arelatively high yield, similar experiments with HCV have not beensuccessful to date (Khromykh et al., 1997, J. Virol. 71, 1497; Behrenset al., 1998, J. Virol. 72, 2364; Moser et al., 1998 J. Virol. 72,5318). It is known from different publications that cell lines orprimary cell cultures can be infected with high-titer patient serumcontaining HCV (Lanford et al. 1994 Virology 202, 606; Shimizu et al.1993 PNAS, USA 90, 6037-6041; Mizutani et al. 1996 J. Virol. 70,7219-7223; Ikda, et al. 1998, Virus Res. 56, 157; Fourner et al. 1998,J. Gen. Virol. 79, 2376; Ito et al. 1996, J. Gen. Virol. 77, 1043-1054).However, these virus-infected cell lines or cell cultures do not allowthe direct detection of HCV-RNA or HCV antigens.

[0010] It is also known from the publications of Yoo et al. 1995 J.Virol., 69, 32-38; and of Dash et al., 1997, Am. J. Pathol., 151,363-373; that hepatoma cell lines can be transfected with syntheticHCV-RNA obtained through in vitro transcription of the cloned HCVgenome. In both publications the authors started from the basic ideathat the viral HCV genome is a plus-strand RNA functioning directly asmRNA after being transfected into the cell, permitting the synthesis ofviral proteins in the course of the translation process, and so new HCVparticles could form HCV viruses and their RNA detected through RT-PCR.However the published results of the RT-PCR experiments indicate thatthe HCV replication in the described HCV transfected hepatoma cells isnot particularly efficient and not sufficient to measure the quality ofreplication, let alone measure the modulations in replication afterexposure to potential antiviral drugs. Furthermore it is now known thatthe highly conserved 3′ NTR is essential for the virus replication(Yanagi et al., 1999 Proc. Natl. Acad. Sci. USA, 96, 2291-95). Thisknowledge strictly contradicts the statements of Yoo et al. J. Virol.,69, 32-38(supra) and Dash et al., 1997, Am. J. Pathol., 151, 363-373.(supra), who used for their experiments only HCV genomes with shorter 3′NTRs and not the authentic 3′ end of the HCV genome.

[0011] In WO 98/39031, Rice et al. disclosed authentic HCV genome RNAsequences, in particular containing: a) the highly conserved 5′-terminalsequence “GCCAGCC”; b) the HCV polyprotein coding region; and c) 3′-NTRauthentic sequences.

[0012] In WO 99/04008, Purcell et al. disclosed an HCV infectious clonethat also contained only the highly conserved 5′-terminal sequence“GCCAGC”. Recently Lohman et al. 1999 (Science 285,110-113) andBartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844(in CA2,303,526, laid-open on Oct. 3, 2000) disclosed a HCV cell culturesystem where the viral RNA (1377/NS2-3′) self-replicates in thetransfected cells with such efficiency that the quality of replicationcan be measured with accuracy and reproducibility. The Lohman andBartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844 disclosureswere the first demonstration of HCV RNA replication in cell culture thatwas substantiated through direct measurement by Northern blots. Thisreplicon system and sequences disclosed therein highlight once again theconserved 5′ sequence “GCCAGC”. A similar observation highlighting theconservation of the 5′NTR was made by Blight et al. 2000 (Science290,1972-1974) and WO 01/89364 published on Nov. 29, 2001.

[0013] In addition to the conservation of the 5′ and 3′ untranslatedregions in cell culture replicating RNAs, three other publications byLohman et al. 2001, J. Virol. 1437-1449 Krieger et al. 2001 J. Virol.4614-4624 and Guo et al., (2001) J. Virol. 8516-8523 have recentlydisclosed distinct adaptive mutants within the HCV non-structuralprotein coding region. Specific nucleotide changes that alter the aminoacids of the HCV non-structural proteins are shown to enhance theefficiency of establishing stable replicating HCV subgenomic repliconsin culture cells.

[0014] Applicant has now found that, contrary to all previous reports,the highly conserved 5′-NTR can be mutated by adaptation to give rise toa HCV RNA sequence that, in conjunction with mutations in the HCVnon-structural region, provides for a greater efficiency of transductionand/or replication.

[0015] Applicant has also identified novel adaptive mutations within theHCV non-structural region that improves the efficiency of establishingpersistently replicating HCV RNA in cell culture.

[0016] One advantage of the present invention is to provide analternative to these existing systems comprising a HCV RNA molecule thatself-replicates. Moreover, the present invention demonstrates that theinitiating nucleotide of the plus-strand genome can be either an A as analternative to the G already disclosed.

[0017] A further advantage of the present invention is to provide aunique HCV RNA molecule that transduces and/or replicates with higherefficiency. The Applicant demonstrates the utility of this specific RNAmolecule in a cell line and its use in evaluating a specific inhibitorof HCV replication.

SUMMARY OF THE INVENTION

[0018] In a first embodiment, the present invention provides a 5′-nontranslated region of the hepatitis C virus wherein its highly conservedguanine at position 1 is substituted for adenine.

[0019] Particularly, the present invention provides a hepatitis C viruspolynucleotide comprising adenine at position 1 as numbered according tothe 1377/NS2-3′ construct (Lohmann et al. 1999, Science 285,110-113,Accession #AJ242651). Particularly, the invention provides a HCVself-replicating polynucleotide comprising a 5′-terminus consisting ofACCAGC (SEQ ID NO. 8).

[0020] In a second embodiment, the present invention is directed to aHCV self-replicating polynucleotide encoding a polyprotein comprisingone or more amino acid substitution selected from the group consistingof: R(1135)K; S(1148)G; S(1560)G; K(1691)R; L(1701)F; 1(1984)V;T(1993)A; G(2042)C; G(2042)R; S(2404)P; L(2155)P; P(2166)L and M(2992)T.

[0021] Particularly, the invention is directed to a HCV self-replicatingpolynucleotide encoding a polyprotein comprising the any one of theamino acid substitutions as described above, further comprising theamino acid substitution E(1202)G.

[0022] More particularly, the invention provides a HCV self-replicatingpolynucleotide encoding a polyprotein comprising a G2042C or a G2042Rmutation.

[0023] Most particularly, the invention provides for HCVself-replicating polynucleotide comprising a nucleotide substitutionG->A at position 1, and said polynucleotide encodes a polyproteinfurther comprising a G2042C or a G2042R mutation.

[0024] Particularly, the polynucleotide of the present invention can bein the form of RNA or DNA that can be transcribed to RNA.

[0025] In a third embodiment, the invention also provides for anexpression vector comprising a DNA form of the above polynucleotide,operably linked with a promoter.

[0026] According to a fourth embodiment, there is provided a host celltransfected with the self-replicating polynucleotide or the vector asdescribed above.

[0027] In a fifth embodiment, the present invention provides a RNAreplication assay comprising the steps of:

[0028] incubating the host cell as described above in the absence orpresence of a potential hepatitis C virus inhibitor;

[0029] isolating the total cellular RNA from the cells;

[0030] analyzing the RNA so as to measure the amount of HCV RNAreplicated;

[0031] comparing the levels of HCV RNA in cells in the absence andpresence of the inhibitor.

[0032] In a sixth embodiment, the invention is directed to a method fortesting a compound for inhibiting HCV replication, including the stepsof:

[0033] a) treating the above described host cell with the compound;

[0034] b) evaluating the treated host cell for reduced replication,wherein reduced replication indicates the ability of the compound toinhibit replication.

DETAILED DESCRIPTION OF THE DRAWINGS

[0035]FIG. 1 is a schematic view of the bi-cistronic replicon RNA. Thesequence deviations between the 1377/NS2-3′ replicon from Lohman et al.,1999 Science 285: 110-113 and the APGK12 replicon are indicated belowthe replicon. In place of a G nucleotide at the +1 position in theI377/NS2-3′ replicon, the APGK12 contains an additional G resulting inGG at the 5′ terminus (the first G being counted as position −1). In thelinker region between the neo gene and the EMCV IRES sequence two areasdeviate from I377/NS2-3′: 14 nucleotides (CGCGCCCAGATGTT) which are notpresent in I377/NS2/3′ are inserted at position 1184 in APGK12; 11nucleotides (1231-1241) present in 1377/NS2-3′ are deleted to generateAPGK-12. In the NS5B coding region, a T at position 8032 was mutated toC to eliminate a Ncol restriction site.

[0036]FIG. 2 shows Northern blots of RNA-transfected Huh-7 cell lines.12 μg of total cellular RNA or control RNA was separated on 0.5%agarose-formaldehyde gels and transferred to Hybond N+ paper, fixed and(FIG. 2A) radioactively probed with HCV specific minus-strand RNA thatdetects the presence of plus-strand replicon RNA. Lanes 1 and 2:positive controls that contain 10⁹ copies of in vitro transcribed APGK12RNA. Lane 3: negative control of total cellular RNA from untransfectedHuh-7 cells. Lanes 4 and 5: cellular RNA from B1 and B3 cell lines thathave integrated DNA copies of the neomycin phosphotransferase gene. Lane6: total cellular RNA from a Huh-7 cell line, designated S22.3, thatharbors high copy number HCV sub-genomic replicon RNA as highlighted bythe arrow. Other cell lines have no detectable replicon RNA. FIG. 2B isidentical to FIG. 2A with the exception that the blot was radioactivelyprobed with HCV specific plus-strand RNA to detect the presence of HCVminus-strand RNA. Lanes 1 and 2 are positive control lanes that contain10⁹ copies of full length HCV minus strand RNA. Lane 6, which contains12 μg of total cellular RNA from cell line S22.3, harbors detectableminus-strand replicon RNA at the expected size of 8-9 kilobases. Mrepresent the migration of non-radioactive molecular size markers on theagarose gel. 28s represents the migration of 28s ribosomal RNA andaccounts for the detection of this species in a samples of totalcellular RNA.

[0037]FIG. 3 shows indirect immunofluorescence of a HCV non-structuralprotein in the S22.3 cell line. Indirect immunofluorescence wasperformed on cells that were cultured and fixed, permeabilized andexposed to a rabbit polyclonal antibody specific for a segment of theHCV NS4A protein. Secondary goat anti-rabbit antibody conjugated withred-fluor Alexa 594 (Molecular Probes) was used for detection. Toppanels shows the results of immunofluorescence (40×objective) and thespecific staining of the S22.3 cells. The bottom panels represent theidentical field of cells viewed by diffractive interference contrast(DIC) microscopy. The majority of S22.3 (FIG. 3A) cells within the fieldstain positively for HCV NS4A protein that localizes in the cytoplasm,whereas the B1 cells (FIG. 3B) that fail to express any HCV proteins,only have background level of staining.

[0038]FIG. 4 shows Western-blots following SDS-PAGE separation of totalproteins extracted from three cell lines: (i) naïve Huh-7 cell line,(ii) neomycin resistant Huh-7 cell line B1, and (iii) the S22.3 cellline. Panels A, B, and C, demonstrate the results of western blotsprobed with rabbit polyclonal antisera specific for neomycinphosphotransferase (NPT), HCV NS3, and HCV NS5B, respectively.Visualization was achieved through autoradiographic detection of achemiluminescent reactive secondary\goat anti-rabbit antibody. Panel Ashows that the S22.3 RNA replicon cell line, expresses the NPT proteinat levels higher than control B1 cells and that the naïve Huh-7 cellline does not produce the NPT protein. Panels B and C show that only theS22.3 cell line produces the mature HCV NS3 and NS5B proteins,respectively. M represents molecular weight (in kilodaltons) ofpre-stained polypeptide markers.

[0039]FIGS. 5A and 5B identify the nucleotide and amino acid sequencesrespectively that differ from the APGK12 sequence in the different HCVbi-cistronic replicons. The S22.3 adapted replicon is a first generationreplicon selected following the transfection of RNA transcribed from theAPGK12 template. R3, R7, R16 are second generation replicons that wereselected following the transfection of RNA isolated from the S22.3 firstgeneration replicon cell line. FIG. 5A: Nucleotide mutations that werecharacterized in each of the adapted replicons are indicated adjacent tothe respective segment of the replicon (IRES, NS3, NS4A, NS5A, andNS5B). FIG. 5B: Amino acid numbers are numbered according to the fulllength HCV poly-protein with the first amino acid in the second cistroncorresponding to amino acid 810 in NS2 of 1377/NS2-3′ construct.

[0040]FIG. 6 depicts the colony formation efficiency of four in vitrotranscribed HCV sub-genomic bi-cistronic replicon RNAs. The APGK12serves as the reference sequence; highlighted are the initiatingnucleotides of the HCV IRES in each of the constructs and the amino aciddifferences (from the APGK12 reference sequence) in the HCVnon-structural region for the two R3-rep. Note that the in vitrotranscribed APGK-12 RNAs that harbor either a 5′G or 5′A form colonieswith the same efficiency (ca. 80 cfu/μg in panels A and B) followingselection with 0.25 mg/ml G418. RNA isolated from the second generationR3 cell line was reverse transcribed into DNA and cloned into thepAPGK12 vector backbone to generate the R3-rep, which was sequenced andfound to encode additional changes that included the L(2155)Psubstitution in the NS5A segment of the HCV polyprotein (compare R3-repsequence with the R3 sequence in tables 2 and 3). Various quantities ofin vitro transcribed R3-rep-5′A RNA, were transfected into naïve Huh-7cells to determine a colony formation efficiency of 1.2×10⁶ cfu/μg ofRNA (panel C). Various quantities of R3-rep-5′G were also transfectedresulting in a colony formation efficiency of 2×10⁶ cfu/μg of RNA (panelD).

[0041]FIG. 7 displays a typical RT-PCR amplification plot (left panel)and the graphical representation of Ct values versus known HCV RNAquantity in a standard curve (right panel). Each of the plotted curvesin the left panel, graph the increment of fluorescence reporter signal(delta-Rn) versus PCR cycle number for a predetermined quantity of HCVreplicon RNA. The Ct value is obtained by determining the point at whichthe fluorescence exceeds an arbitrary value (horizontal line). The rightpanel demonstrates the linear relationship between starting RNA copynumber of the predetermined standards (large black dots) and the Ctvalue. Smaller dots are the Ct values of RNA samples (containing unknownquantity of HCV replicon RNA) from S22.3 cells treated with variousconcentrations of a specific inhibitor of HCV replication.

[0042]FIG. 8 shows the effect of increasing concentration of inhibitor Aon HCV RNA replicon levels in Huh7 cells. S22.3 cells were grown in thepresence of increasing concentrations of inhibitor A starting at 0.5 nMand ranging to 1024 nM. The inhibitor dose-response curve is the resultof 11 concentrations from serial two-fold dilutions (1:1). One controlwell, without any inhibitor, was also included during the course of theexperiment. The cells were incubated for 4 days in a 5% CO₂ incubator at37° C. Total cellular RNA was extracted, quantified by optical density.HCV replicon RNA was evaluated by real time RT-PCR and plotted as genomeequivalents/μg total RNA as a function of inhibitor concentration

DEFINITIONS

[0043] Unless defined otherwise, the scientific and technological termsand nomenclature used herein have the same meaning as commonlyunderstood by a person of ordinary skill to which this inventionpertains. Generally, the procedures for cell culture, infection,molecular biology methods and the like are common methods used in theart. Such standard techniques can be found in reference manuals such asfor example Sambrook et al. (1989) Molecular Cloning—A LaboratoryManual, Cold Spring Harbor Labs and Ausubel et al. (1994).

[0044] Nucleotide sequences are presented herein by single strand, inthe 5′ to 3′ direction, from left to right, using the one letternucleotide symbols as commonly used in the art and in accordance withthe recommendations of the IUPAC-IUB Biochemical Nomenclature Commission(1972) Biochemistry, 11, 1726-1732.

[0045] The present description refers to a number of routinely usedrecombinant DNA (rDNA) technology terms. Nevertheless, definitions ofselected examples of such rDNA terms are provided for clarity andconsistency.

[0046] The term “DNA segment or molecule or sequence”, is used herein,to refer to molecules comprised of the deoxyribonucleotides adenine (A),guanine (G), thymine (T) and/or cytosine (C). These segments, moleculesor sequences can be found in nature or synthetically derived. When readin accordance with the genetic code, these sequences can encode a linearstretch or sequence of amino acids which can be referred to as apolypeptide, protein, protein fragment and the like.

[0047] As used herein, the term “gene” is well known in the art andrelates to a nucleic acid sequence defining a single protein orpolypeptide. The polypeptide can be encoded by a full-length sequence orany portion of the coding sequence, so long as the functional activityof the protein is retained.

[0048] A “structural gene” defines a DNA sequence which is transcribedinto RNA and translated into a protein having a specific structuralfunction that constitute the viral particles. “Structural proteins”defines the HCV proteins incorporated into the virus particles namely,core “C”, E1, E2, and E2-p7.

[0049] “Non-structural proteins”, defines the HCV proteins that are notcomprised in viral particles namely, NS2, NS3, NS4A, NS5A and NS5B.

[0050] “Restriction endonuclease or restriction enzyme” is an enzymethat has the capacity to recognize a specific base sequence (usually 4,5 or 6 base pairs in length) in a DNA molecule, and to cleave the DNAmolecule at every place where this sequence appears. An example of suchan enzyme is EcoRI, which recognizes the base sequence G↓AATTC andcleaves a DNA molecule at this recognition site.

[0051] “Restriction fragments” are DNA molecules produced by thedigestion of DNA with a restriction endonuclease. Any given genome orDNA segment can be digested by a particular restriction endonucleaseinto at least two discrete molecules of restriction fragments.

[0052] “Agarose gel electrophoresis” is an analytical method forfractionating polynucleotide molecules based on their size. The methodis based on the fact that nucleic acid molecules migrate through a gelas through a sieve, whereby the smallest molecule has the greatestmobility and travels the farthest through the gel. The sievingcharacteristics of the gel retards the largest molecules such that,these have the least mobility. The fractionated polynucleotides can bevisualized by staining the gel using methods well known in the art,nucleic acid hybridization or by tagging the fractionated molecules witha detectable label. All these methods are well known in the art,specific methods can be found in Ausubel et al. (supra).

[0053] “Oligonucleotide or oligomer” is a molecule comprised of two ormore deoxyribonucleotides or ribonucleotides, preferably more thanthree. The exact size of the molecule will depend on many factors, whichin turn depend on the ultimate function or use of the oligonucleotide.An oligonucleotide can be derived synthetically, by cloning or byamplification.

[0054] “Sequence amplification” is a method for generating large amountsof a target sequence. In general, one or more amplification primers areannealed to a nucleic acid sequence. Using appropriate enzymes,sequences found adjacent to, or in between the primers are amplified. Anamplification method used herein is the polymerase chain reaction (PCR)and can be used in conjunction with the reverse-transcriptase (RT) toproduce amplified DNA copies of specific RNA sequences. “Amplificationprimer” refers to an oligonucleotide, capable of annealing to a RNA orDNA region adjacent to a target sequence and serving as the initiationprimer for DNA synthesis under suitable conditions well known in theart. The synthesized primer extension product is complementary to thetarget sequence.

[0055] The term “domain” or “region” refers to a specific amino acidsequence that defines either a specific function or structure within aprotein. As an example herein, is the NS3 protease domain comprisedwithin the HCV non-structural polyprotein.

[0056] The terms “plasmid” “vector” or “DNA construct” are commonlyknown in the art and refer to any genetic element, including, but notlimited to, plasmid DNA, phage DNA, viral DNA and the like which canincorporate the oligonucleotide sequences, or sequences of the presentinvention and serve as DNA vehicle into which DNA of the presentinvention can be cloned. Numerous types of vectors exist and are wellknown in the art.

[0057] The terminology “expression vector” defines a vector as describedabove but designed to enable the expression of an inserted sequencefollowing transformation or transfection into a host. The cloned gene(inserted sequence) is usually placed under the control of controlelement sequences such as promoter sequences. Such expression controlsequences will vary depending on whether the vector is designed toexpress the operably linked gene in vitro or in vivo in a prokaryotic oreukaryotic host or both (shuttle vectors) and can additionally containtranscriptional elements such as enhancer elements, terminationsequences, tissue-specificity elements, and/or translational initiationand termination sites.

[0058] A host cell or indicator cell has been “transfected” by exogenousor heterologous DNA (e.g. a DNA construct) or RNA, when such nucleicacid has been introduced inside the cell. The transfecting DNA may ormay not be integrated (covalently linked) into chromosomal DNA making upthe genome of the cell. In prokaryotes, yeast, and mammalian cells forexample, the transfecting/transforming DNA may be maintained on anepisomal element such as a plasmid. With respect to eukaryotic cells, anexample of a stably transfected cell is one in which the transfectingDNA has become integrated into a chromosome and is inherited by daughtercells through chromosome replication. A host cell or indicator cell canbe transfected with RNA. A cell can be stably transfected with RNA ifthe RNA replicates and copies of the RNA segregate to daughter cellsupon cell division. This stability is demonstrated by the ability of theeukaryotic cell to establish cell lines or clones comprised of apopulation of daughter cells containing the transfecting DNA or RNA.Transfection methods are well known in the art (Sambrook et al., 1989,Molecular Cloning—A Laboratory Manual, Cold Spring Harbor Labs; Ausubelet al., 1994, Current Protocols in Molecular Biology, Wiley, N.Y.). Ifthe RNA encodes for a genetic marker that imparts an observablephenotype, such as antibiotic resistance, then the stable transfectionof replicating RNA can be monitored by the acquisition of such phenotypeby the host cell.

[0059] As used herein the term “transduction” refers to the transfer ofa genetic marker to host cells by the stable transfection of areplicating RNA.

[0060] The nucleotide sequences and polypeptides useful to practice theinvention include without being limited thereto, mutants, homologs,subtypes, quasi-species, alleles, and the like. It is understood thatgenerally, the sequences of the present invention encode a polyprotein.It will be clear to a person skilled in the art that the polyprotein ofthe present invention and any variant, derivative or fragment thereof,is auto-processed to an active protease.

[0061] As used herein, the designation “variant” denotes in the contextof this invention a sequence whether a nucleic acid or amino acid, amolecule that retains a biological activity (either functional orstructural) that is substantially similar to that of the originalsequence. This variant may be from the same or different species and maybe a natural variant or be prepared synthetically. Such variants includeamino acid sequences having substitutions, deletions, or additions ofone or more amino acids, provided the biological activity of the proteinis conserved. The same applies to variants of nucleic acid sequenceswhich can have substitutions, deletions, or additions of one or morenucleotides, provided that the biological activity of the sequence isgenerally maintained.

[0062] The term “derivative” is intended to include any of the abovedescribed variants when comprising additional chemical moiety notnormally a part of these molecules. These chemical moieties can havevarying purposes including, improving a molecule's solubility,absorption, biological half life, decreasing toxicity and eliminating ordecreasing undesirable side effects. Furthermore, these moieties can beused for the purpose of labeling, binding, or they may be comprised infusion product(s). Different moieties capable of mediating the abovedescribed effects can be found in Remington's The Science and Practiceof Pharmacy (1995). Methodologies for coupling such moieties to amolecule are well known in the art.

[0063] The term “fragment” refers to any segment of an identified DNA,RNA or amino acid sequence and/or any segment of any of the variants orderivatives described herein above that substantially retains itsbiological activity (functional or structural) as required by thepresent invention.

[0064] The terms “variant”, “derivative”, and “fragment” of the presentinvention refer herein to proteins or nucleic acid molecules which canbe isolated/purified, synthesized chemically or produced throughrecombinant DNA technology. All these methods are well known in the art.As exemplified herein below, the nucleotide sequences and polypeptidesused in the present invention can be modified, for example by in vitromutagenesis.

[0065] As used herein, the term “HCV polyprotein coding region” meansthe portion of a hepatitis C virus that codes for the polyprotein openreading frame (ORF). This ORF may encode proteins that are the same ordifferent than wild-type HCV proteins. The ORF may also encode only someof the functional protein encoded by wild-type polyprotein codingregion. The protein encoded therein may also be from different isolatesof HCV, and non-HCV protein may also be encoded therein.

[0066] As used herein, the abbreviation “NTR” used in the context of apolynucleotide molecule means a non-translated region. The term “UTR”means untranslated region. Both are used interchangeably.

[0067] Preferred Embodiments

[0068] Particularly, the invention provides a HCV self-replicatingpolynucleotide molecule comprising a 5′-terminus consisting of ACCAGC(SEQ ID NO.8).

[0069] According to the first embodiment of this invention, there isparticularly provided a HCV polynucleotide construct comprising:

[0070] a 5′-non translated region (NTR) comprising the sequence ACCAGCat, or proximal to, its 5′-terminus;

[0071] a HCV polyprotein coding region; and

[0072] a 3′-NTR region.

[0073] In a second embodiment, the present invention is directed to aHCV self-replicating polynucleotide encoding a polyprotein comprisingone or more amino acid substitution selected from the group consistingof: R(1135)K; S(1148)G; S(1560)G; K(1691)R; L(1701)F; 1(1984)V;T(1993)A; G(2042)C; G(2042)R; S(2404)P; L(2155)P; P(2166)L and M(2992)T.

[0074] Particularly, the invention is directed to a HCV self-replicatingpolynucleotide encoding a polyprotein comprising the any one of theamino acid substitutions as described above, further comprising theamino acid substitution E(1202)G.

[0075] Alternatively, the first embodiment of the present invention isdirected to HCV self-replicating polynucleotide molecule comprising aG2042C/R mutation.

[0076] According to the second embodiment, the present inventionparticularly provides a HCV polynucleotide construct comprising:

[0077] a 5′-NTR region comprising the sequence ACCAGC at, or proximalto, its 5′-terminus;

[0078] a HCV polyprotein region coding for a HCV polyprotein comprisinga G(2042)C or a G(2042)R mutation; and

[0079] a 3′-NTR region.

[0080] Preferably, the polynucleotide construct of the present inventionis a DNA or RNA molecule. More preferably, the construct is a RNAmolecule. Most preferably, the construct is a DNA molecule.

[0081] More particularly, the first embodiment of this invention isdirected to a RNA molecule encoded by the DNA molecule selected from thegroup consisting of: SEQ ID NO. 2, 4, 5, 6, 7, 24 and 25.

[0082] Most particularly, the invention provides a DNA molecule selectedfrom the group consisting of: SEQ ID NO. 2, 4, 5, 6, 7, 24 and 25.

[0083] In a third embodiment, the invention also is directed to anexpression vector comprising DNA forms of the above polynucleotide,operably linked with a promoter.

[0084] Preferably, the promoter is selected from the group consistingof: T3, T7 and SP6.

[0085] According to a fourth embodiment, there is provided a host celltransfected with the self-replicating polynucleotide or vector asdescribed above. Particularly, the host cell is a eukaryotic cell line.More particularly, the eukaryotic cell line is a hepatic cell line. Mostparticularly, the hepatic cell line is Huh-7.

[0086] In a fifth embodiment, the present invention provides a RNAreplication assay comprising the steps of:

[0087] a) incubating the host cell as described above under conditionssuitable for RNA replication;

[0088] b) isolating the total cellular RNA from the cells; and

[0089] c) analyzing the RNA so as to measure the amount of HCV RNAreplicated.

[0090] Preferably, the analysis of RNA levels in step c) is carried outby amplifying the RNA by real-time RT-PCR analysis using HCV specificprimers so as to measure the amount of HCV RNA replicated.

[0091] Alternatively in this fifth embodiment, the construct comprises areporter gene, and the analysis of RNA levels in step c) is carried outby assessing the level of reporter expressed.

[0092] According to a preferred aspect of the sixth embodiment, theinvention is directed to a method for testing a compound for inhibitingHCV replication, including the steps of:

[0093] a) carrying step a) as described in the above assay, in thepresence or absence of the compound;

[0094] b) isolating the total cellular RNA from the cells; and

[0095] c) analyzing the RNA so as to measure the amount of HCV RNAreplicated.

[0096] d) comparing the levels of HCV RNA in cells in the absence andpresence of the inhibitor,

[0097] wherein reduced RNA levels is indicative of the ability of thecompound to inhibit replication.

[0098] Preferably, the cell line is incubated with the test compound forabout 3-4 days at a temperature of about 37° C.

EXAMPLES Example 1

[0099] Replicon Constructs (APGK-12; FIG. 1)

[0100] pET9a-EMCV was obtained by ligating an oligonucleotide linker 5′gaattccagatggcgcgcccagatgttaaccagatccatggcacactctagagtactgtcgac 3′ (SEQID NO.9) to pET-9a (Novagen) that was cut with EcoRI and SalI to formthe vector pET-9a-mod. This linker contains the following restrictionsites: EcoRI, AscI, HpaI, NcoI, XbaI, ScaI, SalI. The EMCV IRES wasamplified by PCR from the vector pTM1 with primers 5′cggaatcgttaacagaccacaacggfttccctc 3′ (SEQ ID NO.10) and 5′ggcgtacccatggtattatcgtgtttttca 3′ (SEQ ID NO.11) and ligated intopET-9a-mod via EcoRI and NcoI to form pET-9a-EMCV.

[0101] The sequence of HCV NS2 to NS5B followed by the 3′UTR of HCV wasobtained from the replicon construct 1377/NS2-3′ (Lohman et al., 1999Science 285:110-113; accession number: AJ242651) and synthesized byOperon Technologies Inc. with a T to C change at the Ncol site in NS5Bat nucleotide 8032. This sequence was released from an GenOp® vector(Operon Technologies) with NcoI and ScaI and transferred intopET-9a-EMCV to form pET-9a-EMCV-NS2-5B-3′UTR.

[0102] pET-9a-HCV-neo was obtained by amplification of the HCV IRES froma HCV cDNA isolated from patient serum with primers

[0103] 5′ gcatatgaattctaatacgactcactataggccagcccccgaftg 3′ (SEQ IDNO.12) containing a T7 promoter and primer

[0104] 5′ ggcgcgccctftggttfttctftgaggtttaggattcgtgctcat 3′ (SEQ IDNO.13) and amplification of the neomycin phosphotransferase gene fromthe vector pcDNA 3.1 (Invitrogen) with primers

[0105] 5′ aaagggcgcatgattgaacaagatggattgcacgca 3′ (SEQ ID NO.14) and 5′gcatatgttaactcagaagaactcgtcaagaaggcgata 3′ (SEQ ID NO.15). These two PCRfragments were mixed and amplified with primers

[0106] 5′ gcatatgaattctaatacgactcactataggccagcccccgattg 3′ (SEQ IDNO.16) and

[0107] 5′ gcatatgttaactcagaagaactcgtcaagaaggcgata 3′ (SEQ ID NO.15), cutwith Eco RI and HpaI and transferred into pET-9a-mod to formpet-9a-HCV-neo. The EMCV-NS2-5B-3′UTR was released frompET-9a-EMCV-NS2-5B-3′UTR with HpaI and ScaI and transferred intopet-9a-HCV-neo that was cut with HpaI to form pET-9a-APGK12. This insertwas sequenced with specific successive primers using a ABI Prism®BigDye™ Terminator Cycle sequencing kit and analyzed on ABI Prism® 377DNA Sequencer and is shown in SEQ ID NO 1.

[0108] RNA in vitro Transcription

[0109] pET-9a-APGK12 DNA was cut with Scal for expression of thefull-length replicon or with BglII for expression of a truncatednegative control RNA. DNA was analyzed on a 1% agarose gel and purifiedby Phenol/Chloroform extraction. RNA was produced using a T7 Ribomax®kit (Promega) followed by extraction with phenol/chloroform andprecipitation with 7.5 M LiCl₂. RNA was treated with DNAse I for 15 minto remove the DNA template and further purified with an RNeasy® column(Qiagen). RNA integrity was verified on a denaturing formaldehyde 1%agarose gel.

Example 2

[0110] Primary transfection of Huh7 cells and selection of replicon celllines Human hepatoma Huh7 cells (Health Science Research Resources Bank,Osaka, Japan) were grown in 10% FBS/DMEM. Cells were grown to 70%confluency, trypsinized, washed with phosphate buffered saline (PBS) andadjusted to 1×10⁷ cells/ml of PBS. 800 μl of cells were transferred into0.4 cm cuvettes and mixed with 15 μg of replicon RNA. Cells wereelectroporated using 960 μF, 300 volts for ˜18 msec and evenlydistributed into two 15 cm tissue culture plates and incubated in atissue culture incubator for 24 hours. The selection of first and secondgeneration replicon cell lines was with 10% FBS/DMEM medium supplementedwith 1 mg/ml of G418. Cells were selected for 3-5 weeks until colonieswere observed that were isolated and expanded.

[0111] Following the G418 selection and propagation of Huh-7 cellstransfected with APGK12 (SEQ ID NO. 1) RNA, cells that formed a distinctcolony were treated with trypsin and serially passed into larger cultureflasks to establish cell lines. Approximately 10×10⁶ cells wereharvested from each cell line. The cells were lysed and the totalcellular RNA extracted and purified as outlined in Qiagen RNAeasy®preparatory procedures. FIG. 2 shows the analysis of 12 μg of totalcellular RNA from various cell lines as analyzed on a Northern blot of adenaturing agarose-formaldehyde gel.

[0112]FIG. 2A is a Northern blot (radioactively probed with HCV specificminus-strand RNA) that detects the presence of plus-strand replicon RNA.Lanes 1 and 2 are positive controls that contain 10⁹ copies of in vitrotranscribed APGK12 RNA. Lane 2 contains the in vitro transcribed RNAmixed with 12 μg of total cellular from naïve Huh-7 cells. Lane 3 is anegative control of total cellular RNA from untreated Huh-7 cells. Lanes4 and 5 contain cellular RNA from the B1 and B3 G418 resistant celllines that have DNA integrated copies of the neomycin phosphotransferasegene. Lane 6 contains total cellular RNA from a Huh-7 cell line,designated S22.3, that harbors high copy number of HCV sub-genomicreplicon RNA as detected by the positive signal in the 8 kilo-baserange. Other cell lines have no detectable replicon RNA. FIG. 2B is aNorthern blot of a duplicate of the gel presented in 2A with theexception that the blot was radioactively probed with HCV specificplus-strand RNA to detect the presence of HCV minus-strand RNA (lanes 1and 2 are positive control lanes that contain 109 copies of full lengthgenomic HCV minus strand RNA); only lane 6, which contains 12 μg oftotal cellular RNA from cell line S22.3, harbors detectable minus-strandreplicon RNA at the expected size of 8-9 kilobases. An quantitativeestimation of RNA copy number, based on phosphorimager scanning of theNorthern blots, is approximately 6×10⁷ copies of plus-strand/μg of totalRNA, and 6×10⁶ copies of minus strand/μg of total RNA. The presence ofthe plus-strand and minus-strand intermediate confirms that the HCVsub-genomic RNA is actively replicating in the S22.3 cell line.

Example 3

[0113] S22.3 Cell Line Constitutively Expresses HCV Non-StructuralProteins.

[0114] HCV non-structural protein expression was examined in the S22.3cell line. FIG. 3 displays the result of indirect immunofluorescencethat detects the HCV NS4A protein in the S22.3 cell line and not in thereplicon negative B1 cell line (a G418 resistant Huh-7 cell line).Indirect immunofluorescence was performed on cells that were culturedand fixed (with 4% paraformaldehyde) onto Lab-tek chamber slides. Cellswere permeabilized with 0.2% Triton X-100 for 10 minutes followed by a 1hour treatment with 5% milk powder dissolved in phosphate-bufferedsaline (PBS). A rabbit serum containing polyclonal antibody raisedagainst a peptide spanning the HCV NS4A region was the primary antibodyused in detection. Following a 2 hour incubation with the primaryantibody, cells were washed with PBS and a secondary goat anti-rabbitantibody conjugated with red-fluor Alexa® 594 (Molecular Probes) wasadded to cells for 3 hours. Unbound secondary antibody was removed withPBS washes and cells were sealed with a cover slip. FIG. 3 (top panels)shows the results of immunofluorescence as detected by a microscope withspecific fluorescent filtering; the bottom panels represent theidentical field of cells viewed by diffractive interference contrast(DIC) microscopy. The majority of S22.3 (FIG. 3A) cells within the fieldstain positively for HCV NS4A protein that localizes in the cytoplasm,whereas the B1 cells (FIG. 3B) that fail to express any HCV proteins,only have background level of staining. A small proportion of S22.3cells express high levels of intensely stained HCV NS4A.

[0115] Expression of the proteins encoded by the bi-cistronic repliconRNA was also examined on Western-blots following SDS-PAGE separation oftotal proteins extracted from: (i) naïve Huh-7 cell line, (ii) neomycinresistant Huh-7 cell line B1, and (iii) the S22.3 cell line. FIG. 4panels A, B, and C, demonstrate the results of western blots probed withrabbit polyclonal antisera specific for neomycin phosphotransferase(NPT), HCV NS3, and HCV NS5B, respectively. Visualization was achievedthrough autoradiographic detection of a chemiluminescent reactivesecondary HRP-conjugated goat anti-rabbit antibody. FIG. 4 panel A showsthat the S22.3 RNA replicon cell line, expresses the NPT protein atlevels higher than B1 cells (which contain an integrated DNA copy of thenpt gene) and that the naïve Huh-7 cell line does not produce the NPTprotein. FIG. 4 panels B and C show that only the S22.3 cell lineproduces the mature HCV NS3 and NS5B proteins, respectively. The westernblots demonstrate that the S22.3 cell line, which harbors activelyreplicating HCV sub-genomic replicon RNA, maintains replication of theRNA through the high level expression of the HCV non-structuralproteins.

Example 4 Sequence Determination of Adapted Replicons

[0116] Total RNA was extracted from replicon containing Huh7 cells usinga RNeasy Kit (Qiagen). Replicon RNA was reverse transcribed andamplified by PCR using a OneStep RT-PCR kit (Qiagen) and HCV specificprimers (as selected from the full-length sequence disclosed in WO00/66623). Ten distinct RT-PCR products, that covered the entirebi-cistronic replicon in a staggered fashion, were amplified usingoligonucleotide primers. The PCR fragments were sequenced directly withABI Prism® BigDye™ Terminator Cycle PCR Sequencing and analyzed on ABIPrism® 377 DNA Sequencer. To analyze the sequence of the HCV replicon 3′and 5′ ends a RNA ligation/RT-PCR procedure described in Kolykhalov etal. 1996 J. of Virology, 7, p. 3363-3371 was followed. The nucleotidesequence of S22.3 is presented as SEQ ID NO. 2.

Example 5

[0117] Serial Passage of HCV Replicon RNA

[0118] The total cellular RNA from the S22.3 cell line was prepared asdescribed above. HCV Replicon RNA copy number was determined by Taqman®RT-PCR analysis and 20 μg of total S22.3 cellular RNA (containing 1×10⁹copies of HCV RNA) was transfected by electroporation into 8×10⁶ naïveHuh-7 cells. Transfected cells were subsequently cultured in 10 cmtissue culture plates containing DMEM supplemented with 10% fetal calfserum (10% FCS). Media was changed to DMEM (10% FCS) supplemented with 1mg/ml G418 24 hours after transfection and then changed every threedays. Twenty-three visible colonies formed three to four weekspost-transfection and G418 selection. G418 resistant colonies wereexpanded into second generation cell lines that represent the first celllines harboring serially passaged HCV Replicon RNA. Three of these celllines: R3, R7, and R16 were the subject of further analyses. First, theefficiency of transduction by each of the adapted replicons wasdetermined by electroporation of the total cellular RNA (extracted fromthe R3, R7 and R16) into naïve Huh-7 cells; following electroporation,the transduction efficiency was determined as described above, bycounting the visible G418 resistant colonies that arose following 3 to 5weeks of G418 selection (Table 1). Second, the sequence of the seriallypassed adapted replicons was determined from the total cellular RNA thatwas extracted from each of the R3, R7 and R16 replicon cell lines asdescribed in example 4 (SEQ ID NO.4, 5, 6). Using the pAPGK12 as areference sequence (SEQ ID NO.1), the nucleotide changes that wereselected in HCV segment of the adapted replicons are presented in FIG.5A. Some of these nucleotide changes are silent and do not change theencoded amino acid whereas others result in an amino acid substitution.FIG. 5B summarizes the amino acid changes encoded by the adaptedreplicons with the amino acid sequence of pAPGK12 as the reference. Itis important to note that the reference sequence APGK-12 (SEQ ID NO. 1)contains an extra G at the 5′-terminal (5′-GG) that is not maintained inthe replicating RNA of the established cell lines. Also noteworthy isthat, in addition to G->A at nucleotide 1, there is also an adaptedmutation G->C/R at amino acid 2042 (shown as amino acid 1233 in thesequence listing since a.a. 810 of NS2 is numbered as a.a. 1 in SEQ ID)that can be found in all clones analyzed. TABLE 1 Transfection of Huh-7cells RNA Copies of Replicon # Colonies SEQ ID 5 ng APKG12 replicon 1.2× 10⁹   0 in 20 μg total Huh-7 RNA 15 μg APKG12 3 × 10¹² 1 (S22.3) 1replicon RNA 20 μg total: 3 × 10⁹ 23 (3 clones 2 S22.3 cellular RNAanalyzed) R3 cellular RNA 1 × 10⁹ 200 4 R7 cellular RNA 1 × 10⁹ 20 5 R16cellular RNA 3 × 10⁸ 100 6 cloned R3rep RNA 2.3 × 10⁸   2000 7

Example 6

[0119] Construction of APGK12 with 5′ G->A substitution (APGK12-5′A, SEQID NO.24) The pAPGK12 DNA was modified to change the first nucleotide inthe sequence to replace the 5′GG with a 5′A. The change in the pAPGK12was introduced by replacing an EcoRI/AgeI portion of the sequence with aPCR-generated EcoRI/AgeI fragment that includes the mutation. Theoligonucleotides used for the amplification were (SEQ ID. NO. 20):5′-GTG GAC GAA TTC TAA TAC GAC TCA CTA TAA CCA GCC CCC GAT TGG-3′ and(SEQ ID. NO. 21): 5′-GGA ACG CCC GTC GTG GCC AGC CAC GAT-3′ andgenerated a 195 bp DNA fragment that was then digested with EcoRI andAgeI. The resulting 178 bp restriction fragment was used to replace theEcoRI I AgeI fragment in pAPGK12 to generate the pAPGK12-5′A plasmid.

Example 7

[0120] cDNA Cloning of the R3-Replicon (R3REP).

[0121] The cDNA clone of the R3 replicon was produced by RT-PCR of RNAextracted from the R3 cell line. The following two oligonucleotides wereused: (SEQ ID. NO. 22): 5′-GTC GTC TTC TCT GAC ATG GAG AC-3′ and (SEQID. NO. 23): 5′-GAG TTG CTC AGT GGA TTG ATG GGC AGC-3′. The −400 nt PCRfragment, starting within the NS2 coding region and extending to the5′-end of the NS5B coding region, was cloned into the plasmid pCR3.1 byTA cloning (Invitrogen). The SacII/XhoI portion of this R3 sequence wasthen used to replace the SacII/XhoI fragment present in the pAPGK12 andthe pAPGK12-5′A described above. Consequently, two R3 cDNA sequenceswere generated: (I) R3-Rep-5′G with an initiating 5′G (SEQ ID NO.7), andR3-Rep-5′A (SEQ ID NO.25) with an initiating 5′A. Sequencing of the R3rep cDNA identified unique nucleotide changes that differ from theoriginal pAPGK12 sequence (see FIG. 5A); some of these changes aresilent and do not change the encoded amino acid, whereas others doresult in an amino acid change (see FIG. 5B). The differences between R3and the R3-rep reflect the isolation of a unique R3-rep cDNA cloneencoding nucleotide changes that were not observed from the sequencingof the total RNA extracted from the R3 cell line.

Example 8

[0122] Efficiency of Colony Formation with Modified Constructs

[0123] RNA from pAPGK12, pAPGK12-5′A, pR3-Rep and pR3-Rep-5′A wasgenerated by in vitro transcription using the T7 Ribomax® kit (Promega)as described in example 1 above. The reactions containing thepAPGK12-5′A and pR3-Rep-5′A templates were scaled-up 10-fold due to thelimitation of commercial RNA polymerase in initiating transcripts with5′-A. The full length RNAs and control truncated RNA for each clone wereintroduced into 8×10⁶ naïve Huh-7 cells by electroporation as describedin example 2. Replicon RNA was supplemented with total cellular Huh-7carrier RNA to achieve a final 15-20 μg quantity. The cells were thencultured in DMEM medium supplemented with 10% fetal calf serum and 0.25mg/ml G418 in two 150 mm plates. The lower concentration of G418 wassufficient to isolate and select replicon containing cell lines as noneof the transfectants with the control truncated RNA produced anyresistant colonies. In contrast, in vitro transcribed APGK-12 RNAs thatharbor either a 5′G or 5′A form colonies with the same efficiency (ca.80 cfu/μg in FIG. 6 panels A and B) following selection with G418.Various quantities (ranging from 0.1 ng to 1 μg) of the R3-rep-5′A RNA,were transfected into naïve Huh-7 cells to determine a colony formationefficiency of 1.2×10⁶ cfu/μg of RNA (FIG. 6 panel C depicts transfectionwith 1 μg of RNA). Various quantities (ranging from 0.1 ng to 1 μg) ofR3-rep [5′G] were similarly transfected resulting in a colony formationefficiency of 2×10⁶ cfu/μg of RNA (FIG. 6 panel D depicts colonyformation with 1 μg of RNA). Note that, shown for the first time, HCVsubgenomic replicons replicate as efficiently with a 5′ A nucleotide inplace of the 5′G. APGK12 with a 5′A or 5′G RNA have similar transductionefficiencies. Similarly, R3-Rep RNAs with either the 5′A or 5′G bothdisplay the markedly increased transduction efficiency. Notably, theadaptive mutants within the HCV non-structural segment encoded by theR3-Rep provides for a substantial increase in transduction efficiency asdepicted by the dramatic increase in colony forming units per μg oftransfected RNA.

Example 9

[0124] Quantification of HCV Replicon RNA Levels in Cell Lines

[0125] S22.3 cells, or cell lines harboring other adapted replicons,were seeded in DMEM supplemented with 10% FBS, PenStrep and 1 μg/mLGeneticin. At the end of the incubation period the replicon copy numberis evaluated by real-time RT-PCR with the ABI Prism 7700 SequenceDetection System. The TAQMAN® EZ RT-PCR kit provides a system for thedetection and analysis of HCV RNA (as first demonstrated by Martell etal. 1999 J. Clin. Microbiol. 37: 327-332). Direct detection of thereverse transcription polymerase chain reaction (RT-PCR) product with nodownstream processing is accomplished by monitoring the increase influorescence of a dye-labeled DNA probe (FIG. 6). The nucleotidesequence of both primers (adapted from Ruster, B. Zeuzem, S. and Roth,W. K., 1995. Analytical Biochemistry 224:597-600) and probe (adaptedfrom Hohne, M., Roeske, H. and Schreier, E. 1998, Poster Presentation:P297 at the Fifth International Meeting on Hepatitis C Virus and RelatedViruses Molecular Virology and Pathogenesis, Venezia-Lido Italy, June25-28, 1998) located in the 5′-region of the HCV genome are thefollowing:

[0126] HCV Forward Primer:

[0127] 5′ ACG CAG AAA GCG TCT AGC CAT GGC GTT AGT 3′ (SEQ ID NO.17)

[0128] HCV Reverse Primer:

[0129] 5′ TCC CGG GGC ACT CGC MG CAC CCT ATC AGG 3′ (SEQ ID NO.18)

[0130] HCV Probe:

[0131] 5′ FAM-TGG TCT GCG GAA CGG GTG AGT ACA CC-TAMRA 3′ (SEQ ID NO.19)

[0132] FAM: Fluorescence reporter dye.

[0133] TAMRA: Quencher dye.

[0134] Using The TAQMAN® EZ RT-PCR kit, the following reaction was setup: Volume per sample Component (μL) Final Concentration RNase-FreeWater 16 — 5X Taqman EZ Buffer 10 1X Manganese Acetate 25 mM 6  3 mMdATP 10 mM 1.5 300 μM dCTP 10 mM 1.5 300 μM dGTP 10 mM 1.5 300 μM dUTP20 mM 1.5 300 μM HCV Forward Primer 10 μM 1 200 nM HCV Reverse Primer 10μM 1 200 nM HCV Probe 5 μM 2 200 nM rTth DNA Polymerase 2  0.1 U/μL 2.5U/μL AmpErase UNG 1 U/μL 0.5 0.01 U/μL Total Mix 45 —

[0135] To this reaction mix, 5 μL of total RNA extracted from S22.3cells diluted at 10 ng/μL was added, for a total of 50 ng of RNA perreaction. The replicon copy number was evaluated with a standard curvemade from known amounts of replicon copies (supplemented with 50 ng ofwild type Huh-7 RNA) and assayed in an identical reaction mix (FIG. 7).

[0136] Thermal cycler parameters used for the RT-PCR reaction on the ABIPrism 7700 Sequence Detection System were optimized for HCV detection:Temp- Cycle erature (° C.) Time (Minutes) Repeat Reaction Hold 50 2Initial Step Hold 60 30 Reverse Transcription Hold 95 5 UNG Deactivation95 0:15 2 Melt Cycle 60 1 Anneal/Extend 90 0:15 40 Melt Cycle 60 1Anneal/Extend

[0137] Quantification is based on the threshold cycle, where theamplification plot crosses a defined fluorescence threshold. Comparisonof the threshold cycles provides a highly sensitive measure of relativetemplate concentration in different samples. Monitoring during earlycycles, when PCR fidelity is at its highest, provides precise data foraccurate quantification. The relative template concentration can beconverted to RNA copy numbers by employing a standard curve of HCV RNAwith known copy number (FIG. 7).

Example 10

[0138] A Specific HCV NS3 Protease Anti-Viral Compound InhibitsReplication of the HCV replicon in S22.3 cell lines.

[0139] In order to determine the effect of a specific HCV NS3 proteaseanti-viral compound on replicon levels in S22.3 cells, the cells wereseeded in 24 Well Cell Culture Cluster at 5×10⁴ cells per well in 500 μLof DMEM complemented with 10% FBS, PenStrep and 1 μg/mL Geneticin. Cellswere incubated until compound addition in a 5% CO₂ incubator at 37° C.The dose-response curve of the inhibitor displayed 11 concentrationsresulting from serial two-fold dilutions (1:1). The startingconcentration of compound A was 100 nM. One control well (without anycompound) was also included in the course of the experiment. The 24 wellplates were incubated for 4 days in a 5% CO₂ incubator at 37° C.Following a 4 day incubation period, the cells were washed once with PBSand RNA was extracted with the RNeasy® Mini Kit and Qiashredder® fromQiagen. RNA from each well was eluted in 50 uL of H₂O. The RNA wasquantified by optical density at 260 nm on a Cary 1 E UV-VisibleSpectrophotometer. 50 ng of RNA from each well was used to quantify theHCV replicon RNA copy number as detailed in Example 6. The level ofinhibition (% inhibition) of each well containing inhibitor wascalculated with the following equation (CN=HCV Replicon copy number):${\% \cdot {inhibition}} = {( \frac{{{CN} \cdot {control}} - {{CN} \cdot {well}}}{{CN} \cdot {control}} )*100}$

[0140] The calculated % inhibition values were then used to determineIC₅₀, slope factor (n) and maximum inhibition (I_(max)) by thenon-linear regression routine NLIN procedure of

[0141] SAS using the following equation:${\% \cdot {inhibition}} = \frac{I_{\max} \times \lbrack{inhibitor}\rbrack^{n}}{\lbrack{inhibitor}\rbrack^{n} + {IC}_{50}^{n}}$

[0142] Compound A was tested in the assay at least 4 times. The IC₅₀curves were analyzed individually by the SAS nonlinear regressionanalysis. FIG. 8 shows a typical curve and Table 2 shows the individualand average IC₅₀ values of compound A. The average IC₅₀ of compound A inthe replication assay was 1.1 nM. TABLE 2 IC₅₀ of compound A in theS22.3 Cell line Replicon Assay. Compound IC₅₀ (nM) Average IC₅₀ (nM) A1.2 1.2 1.0 0.9 1.1 ± 0.2

[0143] Discussion

[0144] The reproducible and robust ex vivo propagation of hepatitis Cvirus, to levels required for the accurate testing of potentialanti-viral compounds, has not been achieved with any system. As analternative approach to studying the molecular mechanisms of hepatitis Cvirus RNA replication, selectable self-replicating bi-cistronic RNAswere developed (Lohman et al., 1999, Science 285, 110-113;Bartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844 CA 2,303,526).Minimally, these replicons encode for some or all of the non-structuralproteins and also carry a selectable marker such as the neomycinphosphotransferase. Though intracellular steady-state levels of thesesub-genomic replicon RNAs among the selected clones is moderate to high,the frequency of generating G418-resistant colonies upon transfection ofthe consensus RNA described by Lohman et al. or Bartenschlager, R. etal., 1993, J. Virol., 67, 3835-3844 is very low. Less than 100 coloniesare generated when 8 million cells are transfected with 1 μg of in vitrotranscribed bi-cistronic replicon RNA. A low efficiency of colonyformation was first noted by Lohmann et al (1999 et al, Science 285,110-113). Since then, Lohmann et al. (2001) J. Virol. 1437-1449, Blightet al. 2000, Science 290, 1972-1974,and Guo et al., (2001) J. Virol.8516-8523, have isolated sub-genomic RNAs with markedly improvedefficiencies in the colony formation assay. Lohmann et al., 1999 Science285,110-113 originally reported that selection of sub genomic repliconsmay not involve the selection of adaptive mutants as serially passagedRNA did not demonstrate an improved transfection efficiency.Nevertheless, in an effort to characterize the function and fitness ofreplicating HCV RNA, we serially passaged the replicon RNA that wasisolated from the first selected cell-line. Notably, a significantincrease in colony forming efficiency was obtained from this experiment,even though the quantity of replicon RNA was orders of magnitude lowerthan originally used to transfect the in vitro transcribed RNA.Furthermore, a second round serial passage of replicon RNA from thisfirst generation clone into naïve Huh-7 cells provided for yet anotherincrease in colony formation efficiency (Table 1).

[0145] Our analysis of replicating HCV RNAs identified several adaptivemutations that enhance the efficiency of colony formation by up to 4orders of magnitude. Adaptive mutations were found in manynon-structural proteins, as well as in the 5′ non-translated region. Thesubstitution of the 5′-GG doublet for a 5′-A as the inauguratingnucleotide of the HCV 5′-UTR is a variant of the HCV genome that has notbeen previously described, despite the sequencing of innumerablegenotypes and subtypes from across the world. Our original replicon thatcarried a 5′-GG evolved to variants with either a single 5′-A or 5′-G,both of which showed equal transduction efficiency. We describe here thefirst report of a HCV genome that can tolerate and stably maintain a 5′Aextremity. Moreover, we were successful in re-introducing this definedsingle nucleotide substitution into our cDNA clone and generate in vitrotranscribed RNA harboring such an extremity to confirm that a 5′Afunctions as efficiently as a 5′G.

[0146] We have identified adaptive amino acid substitutions in the HCVnon-structural proteins NS3, NS4A and NS5A in the R3 replicon, and asubstitution in NS5B in the R7 clone (see FIG. 5B). These mutations,particularly the combination defined by the R3-rep (SEQ ID NO. 7), whenreconstituted into a cDNA clone and transcribed onto a RNA replicon,result in a significantly enhanced transduction efficiency of up to20,000 fold from the original wild type APGK12 replicon RNA. However,the steady state levels of intracellular replicon RNA were comparablefrom each of the different isolated clones. This result suggests thatthe increase in replication efficiency by the adaptive mutations doesnot result in higher stable intracellular RNA levels due to higher RNAreplication, but rather confers increased permissivity for establishingthe replicon in a greater number of Huh7 cells. Such a phenotype may bemanifested transiently, through an initial increase of the amount of denovo replication, that is required to surpass a defined threshold toestablish persistently replicating RNAs within a population of dividingcells.

[0147] Recently three other groups also identified other distinctadaptive mutants. Lohmann et al. (2000) reported enhanced transductionefficiencies of up to 10,000 fold with mutations in NS3, NS4B, NS5A andNS5B. Blight et al. 2000, Science 290:1972-1974 reported an augmentationof transduction efficiencies up to 20,000 fold with a single mutation inNS5A whereas Guo et al., (2001) J. Virol. 8516-8523 reported increasesin transduction efficiencies of 5,000-10,000 fold with a deletion of asingle amino acid in NS5A. The amino acid substitutions that we describehere have not previously been identified as adaptive mutants thatenhance the efficiency of RNA transfection and/or replication. Oneexception is the mutation of E11202G in NS3 that we found in both the R7and R116 replicons. This adaptation was previously described by Guo etal., (2001) J. Virol. 8516-8523 and Krieger et al (2001) J. Virol.4614-4624. All other adaptive mutations, without exception, describedherein are unpublished.

[0148] The development of selectable subgenomic HCV replicons hasprovided for potential avenues of exploration on HCV RNA replication,persistence, and pathogenesis in cultured cells. However, the lowtransduction efficiency with the HCV RNA-containing replicons asoriginally described (Lohmann et al., 1999 Science 285: 110-113) showedthat it was not a practical system for reverse genetics studies. Theadaptive mutants described herein overcome the low transductionefficiency. In light of the recent descriptions of adaptive mutants byother groups, we note that adaptation can be achieved by distinctmutations in different HCV NS proteins, although the level of adaptationcan vary drastically. The replicons encoding adaptive mutants that aredescribed herein are ideally suited for reverse genetic studies toidentify novel HCV targets or host cell targets that may modulate HCVRNA replication or HCV replicon RNA colony formation. The adapted andhighly efficient replicons are suitable tools for characterizing subtlegenotypic or phenotypic changes that affect an easily quantifiabletransduction efficiency.

[0149] Lastly, we have used our adapted HCV sub genomic repliconcell-line to demonstrate the proficient inhibition of HCV RNAreplication by a specific small molecule inhibitor of the HCV NS3protease. This is the first demonstration that an antiviral, designed tospecifically inhibit one of the HCV non-structural proteins, inhibitsHCV RNA replication in cell culture. Moreover, this compound and ourS22.3 cell line validate the proposal that RNA replication is directedby the HCV non-structural proteins NS3 to NS5B. The assay that we havedescribed and validated will be extremely useful in characterizing otherinhibitors of HCV non-structural protein function in cell culture in ahigh throughput fashion.

[0150] All references found throughout the present disclosure are hereinincorporated by reference whether they be found in the following list ornot.

1 25 1 8639 DNA HCV CDS (1803)...(8408) 1 ggccagcccc cgattgggggcgacactcca ccatagatca ctcccctgtg aggaactact 60 gtcttcacgc agaaagcgtctagccatggc gttagtatga gtgtcgtgca gcctccagga 120 ccccccctcc cgggagagccatagtggtct gcggaaccgg tgagtacacc ggaattgcca 180 ggacgaccgg gtcctttcttggatcaaccc gctcaatgcc tggagatttg ggcgtgcccc 240 cgcgagactg ctagccgagtagtgttgggt cgcgaaaggc cttgtggtac tgcctgatag 300 ggtgcttgcg agtgccccgggaggtctcgt agaccgtgca ccatgagcac gaatcctaaa 360 cctcaaagaa aaaccaaagggcgcgccatg attgaacaag atggattgca cgcaggttct 420 ccggccgctt gggtggagaggctattcggc tatgactggg cacaacagac aatcggctgc 480 tctgatgccg ccgtgttccggctgtcagcg caggggcgcc cggttctttt tgtcaagacc 540 gacctgtccg gtgccctgaatgaactgcag gacgaggcag cgcggctatc gtggctggcc 600 acgacgggcg ttccttgcgcagctgtgctc gacgttgtca ctgaagcggg aagggactgg 660 ctgctattgg gcgaagtgccggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 720 aaagtatcca tcatggctgatgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 780 ccattcgacc accaagcgaaacatcgcatc gagcgagcac gtactcggat ggaagccggt 840 cttgtcgatc aggatgatctggacgaagag catcaggggc tcgcgccagc cgaactgttc 900 gccaggctca aggcgcgcatgcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 960 tgcttgccga atatcatggtggaaaatggc cgcttttctg gattcatcga ctgtggccgg 1020 ctgggtgtgg cggaccgctatcaggacata gcgttggcta cccgtgatat tgctgaagag 1080 cttggcggcg aatgggctgaccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 1140 cagcgcatcg ccttctatcgccttcttgac gagttcttct gagttcgcgc ccagatgtta 1200 acagaccaca acggtttccctctagcggga tcaattccgc ccccccccct aacgttactg 1260 gccgaagccg cttggaataaggccggtgtg cgtttgtcta tatgttattt tccaccatat 1320 tgccgtcttt tggcaatgtgagggcccgga aacctggccc tgtcttcttg acgagcattc 1380 ctaggggtct ttcccctctcgccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag 1440 cagttcctct ggaagcttcttgaagacaaa caacgtctgt agcgaccctt tgcaggcagc 1500 ggaacccccc acctggcgacaggtgcctct gcggccaaaa gccacgtgta taagatacac 1560 ctgcaaaggc ggcacaaccccagtgccacg ttgtgagttg gatagttgtg gaaagagtca 1620 aatggctctc ctcaagcgtattcaacaagg ggctgaagga tgcccagaag gtaccccatt 1680 gtatgggatc tgatctggggcctcggtgca catgctttac atgtgtttag tcgaggttaa 1740 aaaacgtcta ggccccccgaaccacgggga cgtggttttc ctttgaaaaa cacgataata 1800 cc atg gac cgg gag atggca gca tcg tgc gga ggc gcg gtt ttc gta 1847 Met Asp Arg Glu Met Ala AlaSer Cys Gly Gly Ala Val Phe Val 1 5 10 15 ggt ctg ata ctc ttg acc ttgtca ccg cac tat aag ctg ttc ctc gct 1895 Gly Leu Ile Leu Leu Thr Leu SerPro His Tyr Lys Leu Phe Leu Ala 20 25 30 agg ctc ata tgg tgg tta caa tatttt atc acc agg gcc gag gca cac 1943 Arg Leu Ile Trp Trp Leu Gln Tyr PheIle Thr Arg Ala Glu Ala His 35 40 45 ttg caa gtg tgg atc ccc ccc ctc aacgtt cgg ggg ggc cgc gat gcc 1991 Leu Gln Val Trp Ile Pro Pro Leu Asn ValArg Gly Gly Arg Asp Ala 50 55 60 gtc atc ctc ctc acg tgc gcg atc cac ccagag cta atc ttt acc atc 2039 Val Ile Leu Leu Thr Cys Ala Ile His Pro GluLeu Ile Phe Thr Ile 65 70 75 acc aaa atc ttg ctc gcc ata ctc ggt cca ctcatg gtg ctc cag gct 2087 Thr Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu MetVal Leu Gln Ala 80 85 90 95 ggt ata acc aaa gtg ccg tac ttc gtg cgc gcacac ggg ctc att cgt 2135 Gly Ile Thr Lys Val Pro Tyr Phe Val Arg Ala HisGly Leu Ile Arg 100 105 110 gca tgc atg ctg gtg cgg aag gtt gct ggg ggtcat tat gtc caa atg 2183 Ala Cys Met Leu Val Arg Lys Val Ala Gly Gly HisTyr Val Gln Met 115 120 125 gct ctc atg aag ttg gcc gca ctg aca ggt acgtac gtt tat gac cat 2231 Ala Leu Met Lys Leu Ala Ala Leu Thr Gly Thr TyrVal Tyr Asp His 130 135 140 ctc acc cca ctg cgg gac tgg gcc cac gcg ggccta cga gac ctt gcg 2279 Leu Thr Pro Leu Arg Asp Trp Ala His Ala Gly LeuArg Asp Leu Ala 145 150 155 gtg gca gtt gag ccc gtc gtc ttc tct gat atggag acc aag gtt atc 2327 Val Ala Val Glu Pro Val Val Phe Ser Asp Met GluThr Lys Val Ile 160 165 170 175 acc tgg ggg gca gac acc gcg gcg tgt ggggac atc atc ttg ggc ctg 2375 Thr Trp Gly Ala Asp Thr Ala Ala Cys Gly AspIle Ile Leu Gly Leu 180 185 190 ccc gtc tcc gcc cgc agg ggg agg gag atacat ctg gga ccg gca gac 2423 Pro Val Ser Ala Arg Arg Gly Arg Glu Ile HisLeu Gly Pro Ala Asp 195 200 205 agc ctt gaa ggg cag ggg tgg cga ctc ctcgcg cct att acg gcc tac 2471 Ser Leu Glu Gly Gln Gly Trp Arg Leu Leu AlaPro Ile Thr Ala Tyr 210 215 220 tcc caa cag acg cga ggc cta ctt ggc tgcatc atc act agc ctc aca 2519 Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys IleIle Thr Ser Leu Thr 225 230 235 ggc cgg gac agg aac cag gtc gag ggg gaggtc caa gtg gtc tcc acc 2567 Gly Arg Asp Arg Asn Gln Val Glu Gly Glu ValGln Val Val Ser Thr 240 245 250 255 gca aca caa tct ttc ctg gcg acc tgcgtc aat ggc gtg tgt tgg act 2615 Ala Thr Gln Ser Phe Leu Ala Thr Cys ValAsn Gly Val Cys Trp Thr 260 265 270 gtc tat cat ggt gcc ggc tca aag accctt gcc ggc cca aag ggc cca 2663 Val Tyr His Gly Ala Gly Ser Lys Thr LeuAla Gly Pro Lys Gly Pro 275 280 285 atc acc caa atg tac acc aat gtg gaccag gac ctc gtc ggc tgg caa 2711 Ile Thr Gln Met Tyr Thr Asn Val Asp GlnAsp Leu Val Gly Trp Gln 290 295 300 gcg ccc ccc ggg gcg cgt tcc ttg acacca tgc acc tgc ggc agc tcg 2759 Ala Pro Pro Gly Ala Arg Ser Leu Thr ProCys Thr Cys Gly Ser Ser 305 310 315 gac ctt tac ttg gtc acg agg cat gccgat gtc att ccg gtg cgc cgg 2807 Asp Leu Tyr Leu Val Thr Arg His Ala AspVal Ile Pro Val Arg Arg 320 325 330 335 cgg ggc gac agc agg ggg agc ctactc tcc ccc agg ccc gtc tcc tac 2855 Arg Gly Asp Ser Arg Gly Ser Leu LeuSer Pro Arg Pro Val Ser Tyr 340 345 350 ttg aag ggc tct tcg ggc ggt ccactg ctc tgc ccc tcg ggg cac gct 2903 Leu Lys Gly Ser Ser Gly Gly Pro LeuLeu Cys Pro Ser Gly His Ala 355 360 365 gtg ggc atc ttt cgg gct gcc gtgtgc acc cga ggg gtt gcg aag gcg 2951 Val Gly Ile Phe Arg Ala Ala Val CysThr Arg Gly Val Ala Lys Ala 370 375 380 gtg gac ttt gta ccc gtc gag tctatg gaa acc act atg cgg tcc ccg 2999 Val Asp Phe Val Pro Val Glu Ser MetGlu Thr Thr Met Arg Ser Pro 385 390 395 gtc ttc acg gac aac tcg tcc cctccg gcc gta ccg cag aca ttc cag 3047 Val Phe Thr Asp Asn Ser Ser Pro ProAla Val Pro Gln Thr Phe Gln 400 405 410 415 gtg gcc cat cta cac gcc cctact ggt agc ggc aag agc act aag gtg 3095 Val Ala His Leu His Ala Pro ThrGly Ser Gly Lys Ser Thr Lys Val 420 425 430 ccg gct gcg tat gca gcc caaggg tat aag gtg ctt gtc ctg aac ccg 3143 Pro Ala Ala Tyr Ala Ala Gln GlyTyr Lys Val Leu Val Leu Asn Pro 435 440 445 tcc gtc gcc gcc acc cta ggtttc ggg gcg tat atg tct aag gca cat 3191 Ser Val Ala Ala Thr Leu Gly PheGly Ala Tyr Met Ser Lys Ala His 450 455 460 ggt atc gac cct aac atc agaacc ggg gta agg acc atc acc acg ggt 3239 Gly Ile Asp Pro Asn Ile Arg ThrGly Val Arg Thr Ile Thr Thr Gly 465 470 475 gcc ccc atc acg tac tcc acctat ggc aag ttt ctt gcc gac ggt ggt 3287 Ala Pro Ile Thr Tyr Ser Thr TyrGly Lys Phe Leu Ala Asp Gly Gly 480 485 490 495 tgc tct ggg ggc gcc tatgac atc ata ata tgt gat gag tgc cac tca 3335 Cys Ser Gly Gly Ala Tyr AspIle Ile Ile Cys Asp Glu Cys His Ser 500 505 510 act gac tcg acc act atcctg ggc atc ggc aca gtc ctg gac caa gcg 3383 Thr Asp Ser Thr Thr Ile LeuGly Ile Gly Thr Val Leu Asp Gln Ala 515 520 525 gag acg gct gga gcg cgactc gtc gtg ctc gcc acc gct acg cct ccg 3431 Glu Thr Ala Gly Ala Arg LeuVal Val Leu Ala Thr Ala Thr Pro Pro 530 535 540 gga tcg gtc acc gtg ccacat cca aac atc gag gag gtg gct ctg tcc 3479 Gly Ser Val Thr Val Pro HisPro Asn Ile Glu Glu Val Ala Leu Ser 545 550 555 agc act gga gaa atc cccttt tat ggc aaa gcc atc ccc atc gag acc 3527 Ser Thr Gly Glu Ile Pro PheTyr Gly Lys Ala Ile Pro Ile Glu Thr 560 565 570 575 atc aag ggg ggg aggcac ctc att ttc tgc cat tcc aag aag aaa tgt 3575 Ile Lys Gly Gly Arg HisLeu Ile Phe Cys His Ser Lys Lys Lys Cys 580 585 590 gat gag ctc gcc gcgaag ctg tcc ggc ctc gga ctc aat gct gta gca 3623 Asp Glu Leu Ala Ala LysLeu Ser Gly Leu Gly Leu Asn Ala Val Ala 595 600 605 tat tac cgg ggc cttgat gta tcc gtc ata cca act agc gga gac gtc 3671 Tyr Tyr Arg Gly Leu AspVal Ser Val Ile Pro Thr Ser Gly Asp Val 610 615 620 att gtc gta gca acggac gct cta atg acg ggc ttt acc ggc gat ttc 3719 Ile Val Val Ala Thr AspAla Leu Met Thr Gly Phe Thr Gly Asp Phe 625 630 635 gac tca gtg atc gactgc aat aca tgt gtc acc cag aca gtc gac ttc 3767 Asp Ser Val Ile Asp CysAsn Thr Cys Val Thr Gln Thr Val Asp Phe 640 645 650 655 agc ctg gac ccgacc ttc acc att gag acg acg acc gtg cca caa gac 3815 Ser Leu Asp Pro ThrPhe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp 660 665 670 gcg gtg tca cgctcg cag cgg cga ggc agg act ggt agg ggc agg atg 3863 Ala Val Ser Arg SerGln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met 675 680 685 ggc att tac aggttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc 3911 Gly Ile Tyr Arg PheVal Thr Pro Gly Glu Arg Pro Ser Gly Met Phe 690 695 700 gat tcc tcg gttctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac 3959 Asp Ser Ser Val LeuCys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr 705 710 715 gag ctc acg cccgcc gag acc tca gtt agg ttg cgg gct tac cta aac 4007 Glu Leu Thr Pro AlaGlu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn 720 725 730 735 aca cca gggttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag agc 4055 Thr Pro Gly LeuPro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser 740 745 750 gtc ttt acaggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act 4103 Val Phe Thr GlyLeu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr 755 760 765 aag cag gcagga gac aac ttc ccc tac ctg gta gca tac cag gct acg 4151 Lys Gln Ala GlyAsp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr 770 775 780 gtg tgc gccagg gct cag gct cca cct cca tcg tgg gac caa atg tgg 4199 Val Cys Ala ArgAla Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp 785 790 795 aag tgt ctcata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg 4247 Lys Cys Leu IleArg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu 800 805 810 815 ctg tatagg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc 4295 Leu Tyr ArgLeu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro 820 825 830 ata accaaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc 4343 Ile Thr LysTyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val 835 840 845 acg agcacc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg 4391 Thr Ser ThrTrp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala 850 855 860 tat tgcctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg 4439 Tyr Cys LeuThr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu 865 870 875 tcc ggaaag ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag 4487 Ser Gly LysPro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu 880 885 890 895 ttcgat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag 4535 Phe AspGlu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln 900 905 910 ggaatg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg 4583 Gly MetGln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu 915 920 925 caaaca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc 4631 Gln ThrAla Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser 930 935 940 aagtgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc 4679 Lys TrpArg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe 945 950 955 atcagc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac 4727 Ile SerGly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn 960 965 970 975ccc gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg 4775 ProAla Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro 980 985 990ctc acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg 4823 LeuThr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val 995 10001005 gcc gcc caa ctt gct cct ccc agc gct gct tct gct ttc gta ggc gcc4871 Ala Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala1010 1015 1020 ggc atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aaggtg ctt 4919 Gly Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys ValLeu 1025 1030 1035 gtg gat att ttg gca ggt tat gga gca ggg gtg gca ggcgcg ctc gtg 4967 Val Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly AlaLeu Val 1040 1045 1050 1055 gcc ttt aag gtc atg agc ggc gag atg ccc tccacc gag gac ctg gtt 5015 Ala Phe Lys Val Met Ser Gly Glu Met Pro Ser ThrGlu Asp Leu Val 1060 1065 1070 aac cta ctc cct gct atc ctc tcc cct ggcgcc cta gtc gtc ggg gtc 5063 Asn Leu Leu Pro Ala Ile Leu Ser Pro Gly AlaLeu Val Val Gly Val 1075 1080 1085 gtg tgc gca gcg ata ctg cgt cgg cacgtg ggc cca ggg gag ggg gct 5111 Val Cys Ala Ala Ile Leu Arg Arg His ValGly Pro Gly Glu Gly Ala 1090 1095 1100 gtg cag tgg atg aac cgg ctg atagcg ttc gct tcg cgg ggt aac cac 5159 Val Gln Trp Met Asn Arg Leu Ile AlaPhe Ala Ser Arg Gly Asn His 1105 1110 1115 gtc tcc ccc acg cac tat gtgcct gag agc gac gct gca gca cgt gtc 5207 Val Ser Pro Thr His Tyr Val ProGlu Ser Asp Ala Ala Ala Arg Val 1120 1125 1130 1135 act cag atc ctc tctagt ctt acc atc act cag ctg ctg aag agg ctt 5255 Thr Gln Ile Leu Ser SerLeu Thr Ile Thr Gln Leu Leu Lys Arg Leu 1140 1145 1150 cac cag tgg atcaac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg 5303 His Gln Trp Ile AsnGlu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp 1155 1160 1165 cta aga gatgtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag 5351 Leu Arg Asp ValTrp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys 1170 1175 1180 acc tggctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc 5399 Thr Trp LeuGln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe 1185 1190 1195 ttctca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc 5447 Phe SerCys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile 1200 1205 12101215 atg caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa5495 Met Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys1220 1225 1230 aac ggt tcc atg agg atc gtg ggg cct agg acc tgt agt aacacg tgg 5543 Asn Gly Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn ThrTrp 1235 1240 1245 cat gga aca ttc ccc att aac gcg tac acc acg ggc ccctgc acg ccc 5591 His Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro CysThr Pro 1250 1255 1260 tcc ccg gcg cca aat tat tct agg gcg ctg tgg cgggtg gct gct gag 5639 Ser Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg ValAla Ala Glu 1265 1270 1275 gag tac gtg gag gtt acg cgg gtg ggg gat ttccac tac gtg acg ggc 5687 Glu Tyr Val Glu Val Thr Arg Val Gly Asp Phe HisTyr Val Thr Gly 1280 1285 1290 1295 atg acc act gac aac gta aag tgc ccgtgt cag gtt ccg gcc ccc gaa 5735 Met Thr Thr Asp Asn Val Lys Cys Pro CysGln Val Pro Ala Pro Glu 1300 1305 1310 ttc ttc aca gaa gtg gat ggg gtgcgg ttg cac agg tac gct cca gcg 5783 Phe Phe Thr Glu Val Asp Gly Val ArgLeu His Arg Tyr Ala Pro Ala 1315 1320 1325 tgc aaa ccc ctc cta cgg gaggag gtc aca ttc ctg gtc ggg ctc aat 5831 Cys Lys Pro Leu Leu Arg Glu GluVal Thr Phe Leu Val Gly Leu Asn 1330 1335 1340 caa tac ctg gtt ggg tcacag ctc cca tgc gag ccc gaa ccg gac gta 5879 Gln Tyr Leu Val Gly Ser GlnLeu Pro Cys Glu Pro Glu Pro Asp Val 1345 1350 1355 gca gtg ctc act tccatg ctc acc gac ccc tcc cac att acg gcg gag 5927 Ala Val Leu Thr Ser MetLeu Thr Asp Pro Ser His Ile Thr Ala Glu 1360 1365 1370 1375 acg gct aagcgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc 5975 Thr Ala Lys ArgArg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser 1380 1385 1390 tca tcagct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act 6023 Ser Ser AlaSer Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr 1395 1400 1405 acccgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg 6071 Thr ArgHis Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu 1410 1415 1420tgg cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat 6119 TrpArg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn 1425 14301435 aag gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat6167 Lys Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp1440 1445 1450 1455 gag agg gaa gta tcc gtt ccg gcg gag atc ctg cgg aggtcc agg aaa 6215 Glu Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg SerArg Lys 1460 1465 1470 ttc cct cga gcg atg ccc ata tgg gca cgc ccg gattac aac cct cca 6263 Phe Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp TyrAsn Pro Pro 1475 1480 1485 ctg tta gag tcc tgg aag gac ccg gac tac gtccct cca gtg gta cac 6311 Leu Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val ProPro Val Val His 1490 1495 1500 ggg tgt cca ttg ccg cct gcc aag gcc cctccg ata cca cct cca cgg 6359 Gly Cys Pro Leu Pro Pro Ala Lys Ala Pro ProIle Pro Pro Pro Arg 1505 1510 1515 agg aag agg acg gtt gtc ctg tca gaatct acc gtg tct tct gcc ttg 6407 Arg Lys Arg Thr Val Val Leu Ser Glu SerThr Val Ser Ser Ala Leu 1520 1525 1530 1535 gcg gag ctc gcc aca aag accttc ggc agc tcc gaa tcg tcg gcc gtc 6455 Ala Glu Leu Ala Thr Lys Thr PheGly Ser Ser Glu Ser Ser Ala Val 1540 1545 1550 gac agc ggc acg gca acggcc tct cct gac cag ccc tcc gac gac ggc 6503 Asp Ser Gly Thr Ala Thr AlaSer Pro Asp Gln Pro Ser Asp Asp Gly 1555 1560 1565 gac gcg gga tcc gacgtt gag tcg tac tcc tcc atg ccc ccc ctt gag 6551 Asp Ala Gly Ser Asp ValGlu Ser Tyr Ser Ser Met Pro Pro Leu Glu 1570 1575 1580 ggg gag ccg ggggat ccc gat ctc agc gac ggg tct tgg tct acc gta 6599 Gly Glu Pro Gly AspPro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val 1585 1590 1595 agc gag gaggct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca 6647 Ser Glu Glu AlaSer Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr 1600 1605 1610 1615 tggaca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg 6695 Trp ThrGly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu 1620 1625 1630ccc atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc 6743 ProIle Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val 1635 16401645 tat gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc6791 Tyr Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr1650 1655 1660 ttt gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtgctc aag 6839 Phe Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val LeuLys 1665 1670 1675 gag atg aag gcg aag gcg tcc aca gtt aag gct aaa cttcta tcc gtg 6887 Glu Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu LeuSer Val 1680 1685 1690 1695 gag gaa gcc tgt aag ctg acg ccc cca cat tcggcc aga tct aaa ttt 6935 Glu Glu Ala Cys Lys Leu Thr Pro Pro His Ser AlaArg Ser Lys Phe 1700 1705 1710 ggc tat ggg gca aag gac gtc cgg aac ctatcc agc aag gcc gtt aac 6983 Gly Tyr Gly Ala Lys Asp Val Arg Asn Leu SerSer Lys Ala Val Asn 1715 1720 1725 cac atc cgc tcc gtg tgg aag gac ttgctg gaa gac act gag aca cca 7031 His Ile Arg Ser Val Trp Lys Asp Leu LeuGlu Asp Thr Glu Thr Pro 1730 1735 1740 att gac acc acc atc atg gca aaaaat gag gtt ttc tgc gtc caa cca 7079 Ile Asp Thr Thr Ile Met Ala Lys AsnGlu Val Phe Cys Val Gln Pro 1745 1750 1755 gag aag ggg ggc cgc aag ccagct cgc ctt atc gta ttc cca gat ttg 7127 Glu Lys Gly Gly Arg Lys Pro AlaArg Leu Ile Val Phe Pro Asp Leu 1760 1765 1770 1775 ggg gtt cgt gtg tgcgag aaa atg gcc ctt tac gat gtg gtc tcc acc 7175 Gly Val Arg Val Cys GluLys Met Ala Leu Tyr Asp Val Val Ser Thr 1780 1785 1790 ctc cct cag gccgtg atg ggc tct tca tac gga ttc caa tac tct cct 7223 Leu Pro Gln Ala ValMet Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro 1795 1800 1805 gga cag cgggtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc 7271 Gly Gln Arg ValGlu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys 1810 1815 1820 cct atgggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act 7319 Pro Met GlyPhe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr 1825 1830 1835 gagaat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg 7367 Glu AsnAsp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu 1840 1845 18501855 gcc ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac7415 Ala Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr1860 1865 1870 atc ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggctat cgc 7463 Ile Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly TyrArg 1875 1880 1885 cgg tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggtaat acc ctc 7511 Arg Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly AsnThr Leu 1890 1895 1900 aca tgt tac ttg aag gcc gct gcg gcc tgt cga gctgcg aag ctc cag 7559 Thr Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala AlaLys Leu Gln 1905 1910 1915 gac tgc acg atg ctc gta tgc gga gac gac cttgtc gtt atc tgt gaa 7607 Asp Cys Thr Met Leu Val Cys Gly Asp Asp Leu ValVal Ile Cys Glu 1920 1925 1930 1935 agc gcg ggg acc caa gag gac gag gcgagc cta cgg gcc ttc acg gag 7655 Ser Ala Gly Thr Gln Glu Asp Glu Ala SerLeu Arg Ala Phe Thr Glu 1940 1945 1950 gct atg act aga tac tct gcc ccccct ggg gac ccg ccc aaa cca gaa 7703 Ala Met Thr Arg Tyr Ser Ala Pro ProGly Asp Pro Pro Lys Pro Glu 1955 1960 1965 tac gac ttg gag ttg ata acatca tgc tcc tcc aat gtg tca gtc gcg 7751 Tyr Asp Leu Glu Leu Ile Thr SerCys Ser Ser Asn Val Ser Val Ala 1970 1975 1980 cac gat gca tct ggc aaaagg gtg tac tat ctc acc cgt gac ccc acc 7799 His Asp Ala Ser Gly Lys ArgVal Tyr Tyr Leu Thr Arg Asp Pro Thr 1985 1990 1995 acc ccc ctt gcg cgggct gcg tgg gag aca gct aga cac act cca gtc 7847 Thr Pro Leu Ala Arg AlaAla Trp Glu Thr Ala Arg His Thr Pro Val 2000 2005 2010 2015 aat tcc tggcta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca 7895 Asn Ser Trp LeuGly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala 2020 2025 2030 agg atgatc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa 7943 Arg Met IleLeu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu 2035 2040 2045 caactt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc 7991 Gln LeuGlu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser 2050 2055 2060att gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt 8039 IleGlu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu 2065 20702075 agc gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg8087 Ser Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val2080 2085 2090 2095 gct tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cgagtc tgg aga 8135 Ala Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg ValTrp Arg 2100 2105 2110 cat cgg gcc aga agt gtc cgc gct agg cta ctg tcccag ggg ggg agg 8183 His Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser GlnGly Gly Arg 2115 2120 2125 gct gcc act tgt ggc aag tac ctc ttc aac tgggca gta agg acc aag 8231 Ala Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp AlaVal Arg Thr Lys 2130 2135 2140 ctc aaa ctc act cca atc ccg gct gcg tcccag ttg gat tta tcc agc 8279 Leu Lys Leu Thr Pro Ile Pro Ala Ala Ser GlnLeu Asp Leu Ser Ser 2145 2150 2155 tgg ttc gtt gct ggt tac agc ggg ggagac ata tat cac agc ctg tct 8327 Trp Phe Val Ala Gly Tyr Ser Gly Gly AspIle Tyr His Ser Leu Ser 2160 2165 2170 2175 cgt gcc cga ccc cgc tgg ttcatg tgg tgc cta ctc cta ctt tct gta 8375 Arg Ala Arg Pro Arg Trp Phe MetTrp Cys Leu Leu Leu Leu Ser Val 2180 2185 2190 ggg gta ggc atc tat ctactc ccc aac cga tga acggggagct aaacactcca 8428 Gly Val Gly Ile Tyr LeuLeu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttccctt tttttttttctttttttttt tttttttttt 8488 tttttttttt ttttctcctt tttttttcct ctttttttccttttctttcc tttggtggct 8548 ccatcttagc cctagtcacg gctagctgtg aaaggtccgtgagccgcttg actgcagaga 8608 gtgctgatac tggcctctct gcagatcaag t 8639 28642 DNA HCV CDS (1802)...(8407) variation 6268 r = a or g 2 accagcccccgattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgcagaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctcccgggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccgggtcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgctagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcgagtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaaaaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttgggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 480 ctgatgccgccgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccggtgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgttccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattgggcgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccatcatggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgaccaccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatcaggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaaggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaatatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggcggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcgaatgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgccttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaacggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgcttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtcttttggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctttcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctggaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaaccccccacctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcggcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcctcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatctgatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 1740 aaacgtctaggccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gaccgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp ArgGlu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg atactc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile LeuLeu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tggtgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp TrpLeu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atcccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile ProPro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgcgcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys AlaIle His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gccata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala IleLeu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tacttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr PheVal Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aaggtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys ValAla Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gcactg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala LeuThr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgggcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp AlaHis Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtcgtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val ValPhe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac accgcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr AlaAla Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc aggggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg GlyArg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag gggtgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly TrpArg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggccta ctt ggc tgc atc atc act agc ctc aca ggc 2521 Gln Gln Thr Arg Gly LeuLeu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aaccag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn GlnVal Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttcctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe LeuAla Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gccggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala GlySer Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tacacc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr ThrAsn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcgcgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala ArgSer Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttggtc acg agg cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu ValThr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac agcagg ggg agc cta ctc tcc ccc agg ccc gtc tcc tac ttg 2857 Gly Asp Ser ArgGly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tcttcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser SerGly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc tttcgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe ArgAla Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gtaccc gtc gag tct atg gaa acc act atg cgg tcc ccg gtc 3001 Asp Phe Val ProVal Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acggac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr AspAsn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc catcta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His LeuHis Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcgtat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala TyrAla Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gccgcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala AlaThr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gaccct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp ProAsn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 cccatc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro IleThr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tctggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser GlyGly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gactcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp SerThr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acggct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr AlaGly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcggtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser ValThr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 ThrGly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgt gat 3577 LysGly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 GluLeu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 TyrArg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 ValVal Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag agc gtc4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser Val 740745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctgctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac cccata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtcacg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcgtat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttgtcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser865 870 875 880 gga arg ccg gcc atc att ccc gac agg gaa gtc ctt tac cgggag ttc 4489 Gly Xaa Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg GluPhe 885 890 895 gat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaacag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu GlnGly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttgctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu LeuGln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaatcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu SerLys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aatttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn PheIle 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cctggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro GlyAsn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc accagc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr SerPro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg ggatgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly TrpVal Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tct gct ttcgta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe ValGly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc cttggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu GlyLys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggggtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly ValAla Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atgccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met ProSer Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcccct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser ProGly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgtcgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg ArgHis Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cggctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg LeuIle Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acgcac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr HisTyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atcctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile LeuSer Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cagtgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln TrpIle Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag acc 5353 ArgAsp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Thr 1170 11751180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gacggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp GlyIle Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc ggacat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly HisVal Lys Asn 1220 1225 1230 tgt tcc atg agg atc gtg ggg cct agg acc tgtagt aac acg tgg cat 5545 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys SerAsn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acgggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr GlyPro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctgtgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu TrpArg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cgg gtgggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val GlyAsp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aagtgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys CysPro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gatggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp GlyVal Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc ctacgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu ArgGlu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ctg gttggg tca cag ctc cca tgc gag ccc gaa ccg gac gta gca 5881 Tyr Leu Val GlySer Gln Leu Pro Cys Glu Pro Glu Pro Asp Val Ala 1345 1350 1355 1360 gtgctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val LeuThr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 AlaLys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 13851390 tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act acc6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctcctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu LeuTrp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tcagaa aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser GluAsn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caagcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln AlaGlu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctgcgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu ArgArg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgcccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg ProAsp Tyr Asn Pro Pro Leu 1475 1480 1485 ttr gag tcc tgg aag gac ccg gactac gtc cct cca gtg gta cac ggg 6313 Xaa Glu Ser Trp Lys Asp Pro Asp TyrVal Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aaggcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys AlaPro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtcctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val LeuSer Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc acaaag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr LysThr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acggca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr AlaThr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg ggatcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly SerAsp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gagccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu ProGly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 15951600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp1605 1610 1615 aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aagctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys LeuPro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aacttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn LeuVal Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aagaag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys LysVal Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgggac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg AspVal Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aaggct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys AlaLys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc ccacat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro HisSer Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cggaac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg AsnLeu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aaggac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys AspLeu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atggca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met AlaLys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggccgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly ArgLys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgtgtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg ValCys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cctcag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro GlnAla Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 GlnArg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 18151820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgtgac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys AspLeu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gagcgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu ArgLeu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aactgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn CysGly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agctgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser CysGly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgtcga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys ArgAla Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc ggagac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly AspAsp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gacgag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp GluAla Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tctgcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser AlaPro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttgata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu IleThr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tctggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser GlyLys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 cccctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro LeuAla Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 SerTrp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 20252030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tactcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr SerIle 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cacggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His GlyLeu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gagatc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu IleAsn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg cccttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro LeuArg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg ctactg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu LeuSer Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttcaac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe AsnTrp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gctgcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala AlaSer Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tacagc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr SerGly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgctgg ttc atg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg TrpPhe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atctat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile TyrLeu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttccccttttttttttt tttttttttc tttttttttt 8487 tttttttttt tttttttttc tccttttttttcctcttttt ttccttttct ttcctttggt 8547 ggctccatct tagccctagt cacggctagctgtgaaaggt ccgtgagccg cttgactgca 8607 gagagtgctg atactggcct ctctgcagatcaagt 8642 3 2201 PRT HCV VARIANT 882 Xaa is Lys or Arg 3 Met Asp ArgGlu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 Leu IleLeu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 Leu IleTrp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 Gln ValTrp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 Ile LeuLeu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 LysIle Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 IleThr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120125 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130135 140 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val145 150 155 160 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys ValIle Thr 165 170 175 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile LeuGly Leu Pro 180 185 190 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu GlyPro Ala Asp Ser 195 200 205 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala ProIle Thr Ala Tyr Ser 210 215 220 Gln Gln Thr Arg Gly Leu Leu Gly Cys IleIle Thr Ser Leu Thr Gly 225 230 235 240 Arg Asp Arg Asn Gln Val Glu GlyGlu Val Gln Val Val Ser Thr Ala 245 250 255 Thr Gln Ser Phe Leu Ala ThrCys Val Asn Gly Val Cys Trp Thr Val 260 265 270 Tyr His Gly Ala Gly SerLys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 Thr Gln Met Tyr ThrAsn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 Pro Pro Gly AlaArg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 Leu TyrLeu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 GlyAsp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360365 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370375 380 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val385 390 395 400 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr PheGln Val 405 410 415 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser ThrLys Val Pro 420 425 430 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu ValLeu Asn Pro Ser 435 440 445 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr MetSer Lys Ala His Gly 450 455 460 Ile Asp Pro Asn Ile Arg Thr Gly Val ArgThr Ile Thr Thr Gly Ala 465 470 475 480 Pro Ile Thr Tyr Ser Thr Tyr GlyLys Phe Leu Ala Asp Gly Gly Cys 485 490 495 Ser Gly Gly Ala Tyr Asp IleIle Ile Cys Asp Glu Cys His Ser Thr 500 505 510 Asp Ser Thr Thr Ile LeuGly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 Thr Ala Gly Ala ArgLeu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 Ser Val Thr ValPro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 Thr GlyGlu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 LysGly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600605 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610615 620 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp625 630 635 640 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val AspPhe Ser 645 650 655 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val ProGln Asp Ala 660 665 670 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly ArgGly Arg Met Gly 675 680 685 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg ProSer Gly Met Phe Asp 690 695 700 Ser Ser Val Leu Cys Glu Cys Tyr Asp AlaGly Cys Ala Trp Tyr Glu 705 710 715 720 Leu Thr Pro Ala Glu Thr Ser ValArg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 Pro Gly Leu Pro Val Cys GlnAsp His Leu Glu Phe Trp Glu Ser Val 740 745 750 Phe Thr Gly Leu Thr HisIle Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 Gln Ala Gly Asp AsnPhe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 Cys Ala Arg AlaGln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 Cys LeuIle Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 TyrArg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840845 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850855 860 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser865 870 875 880 Gly Xaa Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr ArgGlu Phe 885 890 895 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr IleGlu Gln Gly 900 905 910 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala IleGly Leu Leu Gln 915 920 925 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala ProVal Val Glu Ser Lys 930 935 940 Trp Arg Thr Leu Glu Ala Phe Trp Ala LysHis Met Trp Asn Phe Ile 945 950 955 960 Ser Gly Ile Gln Tyr Leu Ala GlyLeu Ser Thr Leu Pro Gly Asn Pro 965 970 975 Ala Ile Ala Ser Leu Met AlaPhe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 Thr Thr Gln His Thr LeuLeu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 Ala Gln Leu AlaPro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 Ile AlaGly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 10351040 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala1045 1050 1055 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp LeuVal Asn 1060 1065 1070 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu ValVal Gly Val Val 1075 1080 1085 Cys Ala Ala Ile Leu Arg Arg His Val GlyPro Gly Glu Gly Ala Val 1090 1095 1100 Gln Trp Met Asn Arg Leu Ile AlaPhe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 Ser Pro Thr His TyrVal Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 Gln Ile LeuSer Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 GlnTrp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 11601165 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Thr1170 1175 1180 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val ProPhe Phe 1185 1190 1195 1200 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp ArgGly Asp Gly Ile Met 1205 1210 1215 Gln Thr Thr Cys Pro Cys Gly Ala GlnIle Thr Gly His Val Lys Asn 1220 1225 1230 Cys Ser Met Arg Ile Val GlyPro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 Gly Thr Phe Pro IleAsn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 Pro Ala ProAsn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 12851290 1295 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro GluPhe 1300 1305 1310 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr AlaPro Ala Cys 1315 1320 1325 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe LeuVal Gly Leu Asn Gln 1330 1335 1340 Tyr Leu Val Gly Ser Gln Leu Pro CysGlu Pro Glu Pro Asp Val Ala 1345 1350 1355 1360 Val Leu Thr Ser Met LeuThr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 Ala Lys Arg ArgLeu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 Ser AlaSer Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 14101415 1420 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu AsnLys 1425 1430 1435 1440 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln AlaGlu Glu Asp Glu 1445 1450 1455 Arg Glu Val Ser Val Pro Ala Glu Ile LeuArg Arg Ser Arg Lys Phe 1460 1465 1470 Pro Arg Ala Met Pro Ile Trp AlaArg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 Xaa Glu Ser Trp Lys AspPro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 Cys Pro Leu ProPro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 LysArg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 15301535 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp1540 1545 1550 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp AspGly Asp 1555 1560 1565 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met ProPro Leu Glu Gly 1570 1575 1580 Glu Pro Gly Asp Pro Asp Leu Ser Asp GlySer Trp Ser Thr Val Ser 1585 1590 1595 1600 Glu Glu Ala Ser Glu Asp ValVal Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 Thr Gly Ala Leu IleThr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 Ile Asn AlaLeu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 AlaThr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 16551660 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu1665 1670 1675 1680 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu LeuSer Val Glu 1685 1690 1695 Glu Ala Cys Lys Leu Thr Pro Pro His Ser AlaArg Ser Lys Phe Gly 1700 1705 1710 Tyr Gly Ala Lys Asp Val Arg Asn LeuSer Ser Lys Ala Val Asn His 1715 1720 1725 Ile Arg Ser Val Trp Lys AspLeu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 Asp Thr Thr Ile MetAla Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 Lys GlyGly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 17801785 1790 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser ProGly 1795 1800 1805 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala LysLys Cys Pro 1810 1815 1820 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe AspSer Thr Val Thr Glu 1825 1830 1835 1840 Asn Asp Ile Arg Val Glu Glu SerIle Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 Pro Glu Ala Arg Gln AlaIle Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 Gly Gly Pro LeuThr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 Cys ArgAla Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 19051910 1915 1920 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile CysGlu Ser 1925 1930 1935 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg AlaPhe Thr Glu Ala 1940 1945 1950 Met Thr Arg Tyr Ser Ala Pro Pro Gly AspPro Pro Lys Pro Glu Tyr 1955 1960 1965 Asp Leu Glu Leu Ile Thr Ser CysSer Ser Asn Val Ser Val Ala His 1970 1975 1980 Asp Ala Ser Gly Lys ArgVal Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 Pro Leu AlaArg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 SerTrp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 20252030 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln2035 2040 2045 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys TyrSer Ile 2050 2055 2060 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg LeuHis Gly Leu Ser 2065 2070 2075 2080 Ala Phe Ser Leu His Ser Tyr Ser ProGly Glu Ile Asn Arg Val Ala 2085 2090 2095 Ser Cys Leu Arg Lys Leu GlyVal Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 Arg Ala Arg Ser ValArg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 Ala Thr CysGly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 LysLeu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 21502155 2160 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu SerArg 2165 2170 2175 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu LeuSer Val Gly 2180 2185 2190 Val Gly Ile Tyr Leu Leu Pro Asn Arg 2195 22004 8643 DNA HCV CDS (1802)...(8407) 4 accagccccc gattgggggc gacactccaccatagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcgttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctgcggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccgctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtcgcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgtagaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatgattgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggctatgactgggc gcaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgcaggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcaggacgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcgacgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatctcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggcggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcgagcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagcatcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcgaggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggccgcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatagcgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcgtgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacgagttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggatcaattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgcgtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaaacctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaatgcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaacaacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctgcggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgttgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggggctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcacatgctttaca tgtgtttagt cgaggttaga 1740 aaacgtctag gccccccgaa ccacggggacgtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcgtgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser CysGly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccgcac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro HisTyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atcacc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile ThrArg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cggggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg GlyGly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag ctaatc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu IlePhe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atggtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met ValLeu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac gggctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly LeuIle Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tatgtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr ValGln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtttat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val TyrAsp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cgagac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg AspLeu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gagacc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu ThrLys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atcatc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile IleLeu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctggga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu GlyPro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cctatt acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro IleThr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atcact agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile ThrSer Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtccaa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val GlnVal Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aatggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn GlyVal Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gccggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala GlyPro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gacctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp LeuVal Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgcacc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys ThrCys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg aag cat gcc gatgtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Lys His Ala Asp ValIle Pro Val Arg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tccccc cgg ccc gtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser ProArg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctctgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu CysPro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acccga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr ArgGly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaaacc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu ThrThr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccggcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro AlaVal Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggtagc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly SerGly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tataag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr LysVal Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggggcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly AlaTyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggggta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly ValArg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tatggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr GlyLys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atcata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile IleIle Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggcatc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly IleGly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtcgtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val ValLeu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat ccaaac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro AsnIle Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttttat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe TyrGly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctcatt ttc tgc cat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu IlePhe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctgtcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu SerGly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gtatcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val SerVal Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gctcta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala LeuMet Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgcaat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys AsnThr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttcacc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe ThrIle Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cagcgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln ArgArg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtgact cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val ThrPro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgcgag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys GluCys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gccgag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala GluThr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg cccgtc tgc cag gac cat ctg gag ttc tgg gag ggc gtc 4057 Pro Gly Leu Pro ValCys Gln Asp His Leu Glu Phe Trp Glu Gly Val 740 745 750 ttt aca ggc ctcacc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu ThrHis Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gacaac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp AsnPhe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gctcag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala GlnAla Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc atacgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile ArgLeu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctggga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu GlyAla Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tacatc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr IleMet Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgggtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp ValLeu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg acaaca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr ThrGly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga aggccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Arg ProAla Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gagatg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu MetGlu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cagctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln LeuAla Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gccacc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala ThrLys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cggacc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg ThrLeu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agcggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser GlyIle Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcgata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala IleAla Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 accacc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr ThrGln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005gcc caa ctt gct cct ccc agc gct gct tcc gct ttc gta ggc gcc ggc 4873 AlaGln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 10151020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcgctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala LeuVal Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gaggac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu AspLeu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc ctagtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu ValVal Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggccca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly ProGly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttcgct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe AlaSer Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cctgag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro GluSer Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt cttacc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu ThrIle Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gaggac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu AspCys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgggat tgg ata tgc acg gtg ttg act gat ttc aag gcc 5353 Arg Asp Val Trp AspTrp Ile Cys Thr Val Leu Thr Asp Phe Lys Ala 1170 1175 1180 tgg ctc cagtcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln SerLys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tcatgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser CysGln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 GlnThr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 12251230 tgt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat5545 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acgccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr ProSer 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gctgct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala AlaGlu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cga gtg ggg gat ttc cactac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His TyrVal Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt caggtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln ValPro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttgcac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu HisArg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtcaca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val ThrPhe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctccca tgc gag ccc gaa ctg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu ProCys Glu Pro Glu Leu Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atgctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met LeuThr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt aggctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg LeuAla Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agccag ctg tct gcg cct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser GlnLeu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt catgac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His AspSer Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cggcag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg GlnGlu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 14351440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc aggaaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg LysPhe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aaccct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn ProPro Leu 1475 1480 1485 ttg gag tcc tgg aag gac ccg gac tac gtc cct ccagtg gta cac ggg 6313 Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro ValVal His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aag gcc cct ccg atacca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile ProPro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tctacc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser ThrVal Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggcagc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly SerSer Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tctcct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser ProAsp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gagtcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu SerTyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat cccgat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro AspLeu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gctagt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala SerGlu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 acg ggcgcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly AlaLeu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atcaat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile AsnAla Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 AlaThr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 16551660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt ctatcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu SerVal Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc agatct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg SerLys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agcaag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser LysAla Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaagac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu AspThr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gaggtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu ValPhe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gctcgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala ArgLeu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaaatg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys MetAla Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atgggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met GlySer Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gagttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu PheLeu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttcgca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe AlaTyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aatgac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn AspIle Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 ProGlu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 18651870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat accctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr LeuThr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aagctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys LeuGln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtcgtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val ValIle Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc ctacgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu ArgAla Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggggac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly AspPro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgctcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys SerSer Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtgtac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val TyrTyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gctgcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala AlaTrp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggcaac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly AsnIle Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctgatg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu MetThr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaaaaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu LysAla Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gagcca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu ProLeu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 20752080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tggaga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp ArgHis 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag gggggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly GlyArg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gtaagg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val ArgThr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttggat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu AspLeu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gacata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp IleTyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tggtgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp CysLeu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc cccaac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro AsnArg * 2195 2200 ggccaatagg ccatcctgtt ttttcccttt tttttttttt tttttttttttttttttttt 8487 tttttttttt tttttttttt ttttcttttt tcccaatttt tttccttttctttcctttgg 8547 tggctccatc ttagccctag tcacggctag ctgtgaaagg tccgtgagccgcttgactgc 8607 agagagtgct gatactggcc tctctgcaga tcaagt 8643 5 8648 DNAHCV CDS (1802)...(8407) 5 gccagccccc gattgggggc gacactccac catagatcactcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgagtgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggtgagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcctggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggccttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcaccatgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaagatggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggcacaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgcccggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagcgcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcactgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatctcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatacgcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacgtactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggctcgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgtcgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctggattcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctacccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacggtatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctgagttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgccccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctatatgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccctgtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctgttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgtagcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaagccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttggatagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggatgcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttacatgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcctttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggcgcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly AlaVal Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aagctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys LeuPhe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gccgag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala GluAla His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgcgat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg AspAla Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt accatc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr IleThr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc caggct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln AlaGly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgtgca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atggct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac catctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcggtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gttatc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val IleThr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggcctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly LeuPro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gcagac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala AspSer 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcctac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala TyrSer 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctcaca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu ThrGly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtctcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val SerThr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgttgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys TrpThr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aagggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys GlyPro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggctgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly TrpGln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggcagc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly SerSer Asp 305 310 315 320 ctt tac ttg gtc acg agg cat gcc gat gtc att ccggtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro ValArg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc agg cccgtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro ValSer Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcgggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser GlyHis Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cgg ggg gttgcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val AlaLys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gga acc act atgcgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Gly Thr Thr Met ArgSer Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccgcag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro GlnThr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aagagc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys SerThr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg cttgtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu ValLeu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atgtct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met SerLys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg accatc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr IleThr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag tttctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe LeuAla Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgtgat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys AspGlu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc acagtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr ValLeu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gccacc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala ThrAla Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gaggag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu GluVal Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaagcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys AlaIle Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgccat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys HisSer Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctcgga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu GlyLeu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc atacca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile ProThr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acgggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr GlyPhe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgtgtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys ValThr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gagacg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu ThrThr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggcagg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly ArgThr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca ggagaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly GluArg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tatgac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr AspAla Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tcagtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser ValArg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc caggac cat ctg gag ttc tgg gag agc gtc 4057 Pro Gly Leu Pro Val Cys Gln AspHis Leu Glu Phe Trp Glu Ser Val 740 745 750 ttt aca ggc ctc acc cac atagac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile AspAla His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccctac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro TyrLeu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct ccacct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro ProPro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aagcct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys ProThr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gttcaa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val GlnAsn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gcatgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala CysMet Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gtaggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val GlyGly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agcgtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser ValVal Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga aag ccg gcc atcatt ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Lys Pro Ala Ile IlePro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gagtgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu CysAla Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaacaa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu GlnPhe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caagcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln AlaGlu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaagcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu AlaPhe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caatat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln TyrLeu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tcactg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser LeuMet Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa catacc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His ThrLeu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa cttgct cct ccc agc gct gct tct gct ttc gta ggc gcc ggc 4873 Ala Gln Leu AlaPro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gctgga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala GlyAla Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 AspIle Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 10501055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggggtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly ValVal 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gagggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu GlyAla Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cggggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg GlyAsn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gacgct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp AlaAla Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc actcag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr GlnLeu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tccacg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser ThrPro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg gtatgc acg gtg ttg act gat ttc aag acc 5353 Arg Asp Val Trp Asp Trp Val CysThr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 tgg ctc cag tcc aag ctcctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu LeuPro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgtggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg GlyTyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acctgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr CysPro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tccatg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser MetArg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 ggaaca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly ThrPhe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 ProAla Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 12701275 1280 tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggcatg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc cccgaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro GluPhe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gctcca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala ProAla Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtcggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val GlyLeu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag cccgaa ccg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro GluPro Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccctcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro SerHis Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg ggatct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly SerPro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcgccc tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala ProSer Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gacgct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp AlaAsp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggcggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly GlyAsn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta attttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile LeuAsp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaagta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu ValSer Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cctcga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro ArgAla Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 LeuGlu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 14951500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tctgcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser AlaLeu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcgtcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser SerAla Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccctcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro SerAsp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tccatg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser MetPro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gacggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp GlySer Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtcgtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val ValCys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acgcca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr ProCys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agcaac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser AsnSer Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tctcgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser ArgSer Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctgcag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu GlnVal Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atgaag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met LysAla Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 GluAla Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 17051710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag acacca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr ProIle 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtccaa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val GlnPro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gtattc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val PhePro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tacgat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr AspVal Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tacgga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr GlyPhe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aatgct tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn AlaTrp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acccgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr ArgCys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gttgag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val GluGlu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc agacag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg GlnAla Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc cccctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro LeuThr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgcgcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg AlaSer Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgttac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys TyrLeu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 19151920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acggag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr GluAla 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaacca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys ProGlu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtgtca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val SerVal Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acccgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr ArgAsp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag acagct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr AlaArg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atgtat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met TyrAla Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttcttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe PheSer Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gattgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp CysGln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac ctacct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu ProGln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tcactc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser LeuHis Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgcctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys LeuArg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgggcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg AlaArg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 AlaThr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 21352140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agcctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser LeuSer Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc acg tgg tgc cta ctc ctactt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Thr Trp Cys Leu Leu Leu LeuSer Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tgaacggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 21952200 ggccaatagg ccatcctgtt tttttccctt ttttcccttt tttttttttt tttttttttt8487 tttttttttt tttttttttt ttccccccct tttttcccct ttttttttcc ttttctttcc8547 tttggtggct ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg8607 actgcagaga gtgctgatac tggcctctct gcagatcaag t 8648 6 8638 DNA HCVCDS (1802)...(8407) 6 accagccccc gattgggggc gacactccac catagatcactcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgagtgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggtgagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcctggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggccttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcaccatgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaagatggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggcacaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgcccggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagcgcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcactgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatctcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatacgcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacgtactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggctcgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgtcgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctggattcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctacccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacggtatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctgagttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgccccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctatatgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccctgtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctgttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgtagcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaagccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttggatagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggatgcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttacatgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcctttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggcgcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly AlaVal Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aagctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys LeuPhe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gccgag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala GluAla His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgcgat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg AspAla Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt accatc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr IleThr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc caggct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln AlaGly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgtgca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atggct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac catctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcggtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gttatc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val IleThr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggcctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly LeuPro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gcagac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala AspSer 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcctac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala TyrSer 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctcaca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu ThrGly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtctcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val SerThr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgttgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys TrpThr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aagggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys GlyPro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggctgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly TrpGln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggcagc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly SerSer Asp 305 310 315 320 ctt tac ttg gtc acg agg cat gcc gat gtc att ccggtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro ValArg Arg Arg 325 330 335 ggc gac ggc agg ggg agc cta ctc tcc ccc agg cccgtc tcc tac ttg 2857 Gly Asp Gly Arg Gly Ser Leu Leu Ser Pro Arg Pro ValSer Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcgggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser GlyHis Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gttgcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val AlaLys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gga acc act atgcgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Gly Thr Thr Met ArgSer Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccgcag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro GlnThr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aagagc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys SerThr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg cttgtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu ValLeu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atgtct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met SerLys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg accatc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr IleThr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag tttctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe LeuAla Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgtgat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys AspGlu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc acagtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr ValLeu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gccacc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala ThrAla Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gaggag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu GluVal Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaagcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys AlaIle Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgccat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys HisSer Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctcgga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu GlyLeu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc atacca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile ProThr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acgggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr GlyPhe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgtgtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys ValThr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gagacg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu ThrThr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggcagg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly ArgThr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca ggagaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly GluArg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tatgac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr AspAla Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tcagtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser ValArg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc caggac cat ctg gag ttc tgg gag agc gtc 4057 Pro Gly Leu Pro Val Cys Gln AspHis Leu Glu Phe Trp Glu Ser Val 740 745 750 ttt aca ggc ctc acc cac atagac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile AspAla His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccctac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro TyrLeu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct ccacct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro ProPro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aagcct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys ProThr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gttcaa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val GlnAsn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gcatgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala CysMet Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gtaggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val GlyGly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agcgtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser ValVal Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga aag ccg gcc atcatt ccc gac agg gaa gtc ttt tac cgg gag ttc 4489 Gly Lys Pro Ala Ile IlePro Asp Arg Glu Val Phe Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gagtgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu CysAla Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaacaa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu GlnPhe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caagcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln AlaGlu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaagcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu AlaPhe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caatat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln TyrLeu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tcactg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser LeuMet Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa catacc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His ThrLeu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa cttgct cct ccc agc gct gct tct gct ttc gta ggc gcc ggc 4873 Ala Gln Leu AlaPro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gctgga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala GlyAla Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 AspIle Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 10501055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggggtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly ValVal 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gagggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu GlyAla Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cggggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg GlyAsn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gacgct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp AlaAla Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc actcag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr GlnLeu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tccacg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser ThrPro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg atatgc acg gtg ttg act gat ttc aag acc 5353 Arg Asp Val Trp Asp Trp Ile CysThr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 tgg ctc cag tcc aag ctcctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu LeuPro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgtggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg GlyTyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acctgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr CysPro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 cgt tccatg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Arg Ser MetArg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 ggaaca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly ThrPhe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 ProAla Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 12701275 1280 tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggcatg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc cccgaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro GluPhe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gctcca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala ProAla Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtcggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val GlyLeu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag cccgaa ccg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro GluPro Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccctcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro SerHis Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg ggatct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly SerPro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcgcct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala ProSer Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gacgct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp AlaAsp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggcggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly GlyAsn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta attttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile LeuAsp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaagta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu ValSer Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cctcga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro ArgAla Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 LeuGlu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 14951500 tgt cca ctg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tctgcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser AlaLeu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcgtcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser SerAla Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccctcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro SerAsp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tccatg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser MetPro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gacggg cct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp GlyPro Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtcgtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val ValCys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acgcca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr ProCys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agcaac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser AsnSer Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tctcgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser ArgSer Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctgcag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu GlnVal Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atgaag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met LysAla Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 GluAla Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 17051710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag acacca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr ProIle 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtccaa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val GlnPro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gtattc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val PhePro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tacgat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr AspVal Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tacgga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr GlyPhe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aatgcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn AlaTrp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acccgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr ArgCys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gttgag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val GluGlu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc agacag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg GlnAla Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc cccctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro LeuThr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgcgcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg AlaSer Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgttac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys TyrLeu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 19151920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acggag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr GluAla 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaacca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys ProGlu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtgtca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val SerVal Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acccgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr ArgAsp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag acagct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr AlaArg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atgtat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met TyrAla Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttcttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe PheSer Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gattgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp CysGln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac ctacct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu ProGln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tcactc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser LeuHis Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgcctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys LeuArg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgggcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg AlaArg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 AlaThr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 21352140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agcctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser LeuSer Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc ctactt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu LeuSer Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tgaacggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 21952200 ggccaatagg ccatcctgtt tttttttttt tttttttttt tttttttttt tttttttttt8487 tttttttttt tttttttttt ttttttcctc ttttttttcc ttttctttcc tttggtggct8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga8607 gtgctgatac tggcctctct gcagatcaag t 8638 7 8638 DNA HCV CDS(1802)...(8407) 7 gccagccccc gattgggggc gacactccac catagatcac tcccctgtgaggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcagcctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccggaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgggcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtactgcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacgaatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcacgcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagacaatcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttctttttgtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcgtggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcgggaagggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgctcctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccggctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatggaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagccgaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccatggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgactgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatattgctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgctcccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcccagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc cccccccctaacgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttattttccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttgacgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcgtgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgaccctttgcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtataagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtggaaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaaggtaccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagtcgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaacacgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gttttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val PheVal Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttcctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe LeuAla Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gcacac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala HisLeu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gccgtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 6570 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 8590 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atcacc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctgccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gacagc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tactcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc acc agc ctc acaggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tccacc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser ThrAla 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tggact gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp ThrVal 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggccca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly ProIle 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tggcaa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp GlnAla 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agctcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser SerAsp 305 310 315 320 ctt tac ttg gtc acg aag cat gcc gat gtc att ccg gtgcgc cgg cgg 2809 Leu Tyr Leu Val Thr Lys His Ala Asp Val Ile Pro Val ArgArg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc cgg ccc gtctcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val SerTyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg gggcac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly HisAla Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcgaag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala LysAla Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atg cggtcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg SerPro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cagaca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln ThrPhe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agcact aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser ThrLys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtcctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val LeuAsn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tctaag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser LysAla His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atcacc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile ThrThr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt cttgcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu AlaAsp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gatgag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp GluCys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtcctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val LeuAsp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc accgct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr AlaThr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gaggtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu ValAla Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gccatc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala IlePro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cattcc aag aag aaa tgc gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His SerLys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc ggactc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly LeuAsn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata ccaact agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro ThrSer Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggcttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly PheThr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtcacc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val ThrGln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acgacg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr ThrThr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc aggact ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg ThrGly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaacgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu ArgPro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gacgcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp AlaGly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gttagg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val ArgLeu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gaccat ctg gag ttc tgg gag ggc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp HisLeu Glu Phe Trp Glu Gly Val 740 745 750 ttt aca ggc ctc acc cac ata gacgcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp AlaHis Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tacctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr LeuVal Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cctcca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro ProSer Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cctacg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro ThrLeu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caaaac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln AsnGlu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgcatg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys MetSer Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggcgga gtc cta gca gct ctg gct gcg tat 4393 Ser Thr Trp Val Leu Val Gly GlyVal Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtggtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val ValIle Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga agg ccg gcc atc attccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Arg Pro Ala Ile Ile ProAsp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgtgcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys AlaSer His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caattc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln PheLys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcggag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala GluAla Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gccttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala PheTrp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tattta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr LeuAla Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctgatg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu MetAla Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat accctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr LeuLeu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gctcct ccc agc gct gct tcc gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala ProPro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct ggagcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly AlaAla Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gatatt ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp IleLeu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 PheLys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 10651070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggggct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly AlaVal 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggtaac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly AsnHis Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gctgca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala AlaAla Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cagctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln LeuLeu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acgcca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr ProCys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg ata tgcacg gtg ttg act gat ttc aag gcc 5353 Arg Asp Val Trp Asp Trp Ile Cys ThrVal Leu Thr Asp Phe Lys Ala 1170 1175 1180 tgg ctc cag tcc aag ctc ctgccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu ProArg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt gggtac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly TyrLys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgccca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys ProCys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tcc atgagg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser Met ArgIle Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga acattc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr PhePro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccggcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro AlaPro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 12751280 tac gtg gag gtt acg cga gtg ggg gat ttc cac tac gtg acg ggc atg5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc cccgaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro GluPhe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gctcca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala ProAla Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtcggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val GlyLeu Asn Gln 1330 1335 1340 tac ccg gtt ggg tca cag ctc cca tgc gag cccgaa ctg gac gta gca 5881 Tyr Pro Val Gly Ser Gln Leu Pro Cys Glu Pro GluLeu Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccctcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro SerHis Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg ggatct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly SerPro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcgcct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala ProSer Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gacgct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp AlaAsp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggcggg aac atc acc cgc gtg gag tca gag aat aag 6121 Arg Gln Glu Met Gly GlyAsn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta attttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile LeuAsp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaagta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu ValSer Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cctcga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro ArgAla Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 LeuGlu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 14951500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tctgcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser AlaLeu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcgtcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser SerAla Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccctcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro SerAsp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tccatg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser MetPro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gacggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp GlySer Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtcgtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val ValCys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acgcca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr ProCys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agcaac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser AsnSer Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tctcgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser ArgSer Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctgcag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu GlnVal Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atgaag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met LysAla Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 GluAla Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 17051710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag acacca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr ProIle 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtccaa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val GlnPro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gtattc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val PhePro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tacgat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr AspVal Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tacgga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr GlyPhe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aatgcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn AlaTrp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acccgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr ArgCys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gttgag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val GluGlu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc agacag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg GlnAla Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc cccctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro LeuThr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgcgcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg AlaSer Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgttac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys TyrLeu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 19151920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acggag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr GluAla 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaacca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys ProGlu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtgtca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val SerVal Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acccgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr ArgAsp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag acagct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr AlaArg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atgtat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met TyrAla Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttcttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe PheSer Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gattgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp CysGln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac ctacct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu ProGln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tcactc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser LeuHis Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgcctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys LeuArg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgggcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg AlaArg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 AlaThr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 21352140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agcctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser LeuSer Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc ctactt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu LeuSer Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tgaacggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 21952200 ggccaatagg ccatcctgtt tttttccctt tttttttttc tttttttttt tttttttttt8487 tttttttttt ttttctcctt tttttttcct ctttttttcc ttttctttcc tttggtggct8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga8607 gtgctgatac tggcctctct gcagatcaag t 8638 8 6 DNA HCV 8 accagc 6 9 63DNA HCV 9 gaattccaga tggcgcgccc agatgttaac cagatccatg gcacactctagagtactgtc 60 gac 63 10 33 DNA HCV 10 cggaatcgtt aacagaccac aacggtttccctc 33 11 30 DNA HCV 11 ggcgtaccca tggtattatc gtgtttttca 30 12 45 DNAHCV 12 gcatatgaat tctaatacga ctcactatag gccagccccc gattg 45 13 45 DNAHCV 13 ggcgcgccct ttggtttttc tttgaggttt aggattcgtg ctcat 45 14 36 DNAHCV 14 aaagggcgca tgattgaaca agatggattg cacgca 36 15 39 DNA HCV 15gcatatgtta actcagaaga actcgtcaag aaggcgata 39 16 45 DNA HCV 16gcatatgaat tctaatacga ctcactatag gccagccccc gattg 45 17 30 DNA HCV 17acgcagaaag cgtctagcca tggcgttagt 30 18 30 DNA HCV 18 tcccggggcactcgcaagca ccctatcagg 30 19 26 DNA HCV Label with FAM fluorescencereporter dye 19 tggtctgcgg aacgggtgag tacacc 26 20 45 DNA HCV 20gtggacgaat tctaatacga ctcactataa ccagcccccg attgg 45 21 27 DNA HCV 21ggaacgcccg tcgtggccag ccacgat 27 22 23 DNA HCV 22 gtcgtcttct ctgacatggagac 23 23 27 DNA HCV 23 gagttgctca gtggattgat gggcagc 27 24 8638 DNA HCVCDS (1802)...(8407) 24 accagccccc gattgggggc gacactccac catagatcactcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgagtgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggtgagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcctggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggccttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcaccatgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaagatggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggcacaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgcccggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagcgcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcactgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatctcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatacgcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacgtactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggctcgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgtcgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctggattcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctacccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacggtatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctgagttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgccccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctatatgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccctgtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctgttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgtagcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaagccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttggatagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggatgcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttacatgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcctttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggcgcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly AlaVal Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aagctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys LeuPhe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gccgag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala GluAla His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgcgat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg AspAla Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt accatc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr IleThr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc caggct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln AlaGly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgtgca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atggct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac catctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcggtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gttatc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val IleThr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggcctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly LeuPro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gcagac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala AspSer 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcctac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala TyrSer 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctcaca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu ThrGly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtctcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val SerThr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgttgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys TrpThr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aagggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys GlyPro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggctgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly TrpGln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggcagc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly SerSer Asp 305 310 315 320 ctt tac ttg gtc acg agg cat gcc gat gtc att ccggtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro ValArg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc agg cccgtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro ValSer Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcgggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser GlyHis Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gttgcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val AlaLys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atgcgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met ArgSer Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccgcag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro GlnThr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aagagc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys SerThr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg cttgtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu ValLeu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atgtct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met SerLys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg accatc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr IleThr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag tttctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe LeuAla Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgtgat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys AspGlu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc acagtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr ValLeu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gccacc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala ThrAla Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gaggag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu GluVal Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaagcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys AlaIle Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgccat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys HisSer Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctcgga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu GlyLeu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc atacca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile ProThr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acgggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr GlyPhe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgtgtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys ValThr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gagacg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu ThrThr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggcagg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly ArgThr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca ggagaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly GluArg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tatgac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr AspAla Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tcagtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser ValArg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc caggac cat ctg gag ttc tgg gag agc gtc 4057 Pro Gly Leu Pro Val Cys Gln AspHis Leu Glu Phe Trp Glu Ser Val 740 745 750 ttt aca ggc ctc acc cac atagac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile AspAla His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccctac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro TyrLeu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct ccacct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro ProPro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aagcct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys ProThr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gttcaa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val GlnAsn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gcatgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala CysMet Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gtaggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val GlyGly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agcgtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser ValVal Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga aag ccg gcc atcatt ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Lys Pro Ala Ile IlePro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gagtgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu CysAla Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaacaa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu GlnPhe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caagcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln AlaGlu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaagcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu AlaPhe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caatat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln TyrLeu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tcactg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser LeuMet Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa catacc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His ThrLeu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa cttgct cct ccc agc gct gct tct gct ttc gta ggc gcc ggc 4873 Ala Gln Leu AlaPro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gctgga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala GlyAla Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 AspIle Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 10501055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggggtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly ValVal 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gagggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu GlyAla Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cggggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg GlyAsn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gacgct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp AlaAla Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc actcag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr GlnLeu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tccacg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser ThrPro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg atatgc acg gtg ttg act gat ttc aag acc 5353 Arg Asp Val Trp Asp Trp Ile CysThr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 tgg ctc cag tcc aag ctcctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu LeuPro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgtggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg GlyTyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acctgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr CysPro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 ggt tccatg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Gly Ser MetArg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 ggaaca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly ThrPhe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 ProAla Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 12701275 1280 tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggcatg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc cccgaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro GluPhe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gctcca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala ProAla Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtcggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val GlyLeu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag cccgaa ccg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro GluPro Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccctcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro SerHis Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg ggatct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly SerPro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcgcct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala ProSer Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gacgct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp AlaAsp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggcggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly GlyAsn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta attttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile LeuAsp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaagta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu ValSer Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cctcga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro ArgAla Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 LeuGlu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 14951500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tctgcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser AlaLeu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcgtcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser SerAla Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccctcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro SerAsp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tccatg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser MetPro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gacggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp GlySer Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtcgtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val ValCys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acgcca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr ProCys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agcaac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser AsnSer Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tctcgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser ArgSer Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctgcag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu GlnVal Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atgaag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met LysAla Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 GluAla Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 17051710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag acacca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr ProIle 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtccaa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val GlnPro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gtattc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val PhePro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tacgat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr AspVal Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tacgga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr GlyPhe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aatgcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn AlaTrp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acccgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr ArgCys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gttgag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val GluGlu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc agacag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg GlnAla Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc cccctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro LeuThr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgcgcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg AlaSer Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgttac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys TyrLeu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 19151920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acggag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr GluAla 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaacca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys ProGlu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtgtca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val SerVal Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acccgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr ArgAsp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag acagct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr AlaArg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atgtat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met TyrAla Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttcttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe PheSer Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gattgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp CysGln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac ctacct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu ProGln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tcactc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser LeuHis Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgcctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys LeuArg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgggcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg AlaArg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 AlaThr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 21352140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agcctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser LeuSer Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc ctactt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu LeuSer Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tgaacggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 21952200 ggccaatagg ccatcctgtt tttttccctt tttttttttc tttttttttt tttttttttt8487 tttttttttt ttttctcctt tttttttcct ctttttttcc ttttctttcc tttggtggct8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga8607 gtgctgatac tggcctctct gcagatcaag t 8638 25 8638 DNA HCV CDS(1802)...(8407) 25 accagccccc gattgggggc gacactccac catagatcactcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgagtgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggtgagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcctggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggccttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcaccatgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaagatggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggcacaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgcccggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagcgcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcactgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatctcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatacgcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacgtactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggctcgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgtcgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctggattcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctacccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacggtatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctgagttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgccccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctatatgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccctgtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctgttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgtagcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaagccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttggatagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggatgcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttacatgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcctttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggcgcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly AlaVal Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aagctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys LeuPhe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gccgag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala GluAla His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgcgat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg AspAla Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt accatc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr IleThr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc caggct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln AlaGly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgtgca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atggct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac catctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcggtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gttatc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val IleThr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggcctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly LeuPro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gcagac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala AspSer 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcctac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala TyrSer 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc acc agc ctcaca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu ThrGly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtctcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val SerThr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgttgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys TrpThr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aagggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys GlyPro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggctgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly TrpGln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggcagc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly SerSer Asp 305 310 315 320 ctt tac ttg gtc acg aag cat gcc gat gtc att ccggtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Lys His Ala Asp Val Ile Pro ValArg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc cgg cccgtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro ValSer Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcgggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser GlyHis Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gttgcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val AlaLys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atgcgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met ArgSer Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccgcag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro GlnThr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aagagc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys SerThr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg cttgtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu ValLeu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atgtct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met SerLys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg accatc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr IleThr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag tttctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe LeuAla Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgtgat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys AspGlu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc acagtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr ValLeu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gccacc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala ThrAla Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gaggag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu GluVal Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaagcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys AlaIle Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgccat tcc aag aag aaa tgc gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys HisSer Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctcgga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu GlyLeu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc atacca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile ProThr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acgggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr GlyPhe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgtgtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys ValThr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gagacg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu ThrThr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggcagg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly ArgThr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca ggagaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly GluArg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tatgac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr AspAla Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tcagtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser ValArg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc caggac cat ctg gag ttc tgg gag ggc gtc 4057 Pro Gly Leu Pro Val Cys Gln AspHis Leu Glu Phe Trp Glu Gly Val 740 745 750 ttt aca ggc ctc acc cac atagac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile AspAla His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccctac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro TyrLeu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct ccacct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro ProPro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aagcct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys ProThr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gttcaa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val GlnAsn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gcatgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala CysMet Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gtaggc gga gtc cta gca gct ctg gct gcg tat 4393 Ser Thr Trp Val Leu Val GlyGly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agcgtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser ValVal Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga agg ccg gcc atcatt ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Arg Pro Ala Ile IlePro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gagtgt gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu CysAla Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaacaa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu GlnPhe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caagcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln AlaGlu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaagcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu AlaPhe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caatat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln TyrLeu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tcactg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser LeuMet Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa catacc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His ThrLeu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa cttgct cct ccc agc gct gct tcc gct ttc gta ggc gcc ggc 4873 Ala Gln Leu AlaPro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gctgga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala GlyAla Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 AspIle Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 10501055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggggtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly ValVal 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gagggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu GlyAla Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cggggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg GlyAsn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gacgct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp AlaAla Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc actcag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr GlnLeu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tccacg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser ThrPro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg atatgc acg gtg ttg act gat ttc aag gcc 5353 Arg Asp Val Trp Asp Trp Ile CysThr Val Leu Thr Asp Phe Lys Ala 1170 1175 1180 tgg ctc cag tcc aag ctcctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu LeuPro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgtggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg GlyTyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acctgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr CysPro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tccatg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser MetArg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 ggaaca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly ThrPhe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 ProAla Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 12701275 1280 tac gtg gag gtt acg cga gtg ggg gat ttc cac tac gtg acg ggcatg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc cccgaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro GluPhe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gctcca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala ProAla Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtcggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val GlyLeu Asn Gln 1330 1335 1340 tac ccg gtt ggg tca cag ctc cca tgc gag cccgaa ctg gac gta gca 5881 Tyr Pro Val Gly Ser Gln Leu Pro Cys Glu Pro GluLeu Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccctcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro SerHis Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg ggatct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly SerPro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcgcct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala ProSer Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gacgct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp AlaAsp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggcggg aac atc acc cgc gtg gag tca gag aat aag 6121 Arg Gln Glu Met Gly GlyAsn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta attttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile LeuAsp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaagta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu ValSer Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cctcga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro ArgAla Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 LeuGlu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 14951500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tctgcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser AlaLeu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcgtcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser SerAla Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccctcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro SerAsp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tccatg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser MetPro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gacggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp GlySer Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtcgtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val ValCys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acgcca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr ProCys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agcaac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser AsnSer Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tctcgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser ArgSer Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctgcag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu GlnVal Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atgaag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met LysAla Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 GluAla Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 17051710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag acacca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr ProIle 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtccaa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val GlnPro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gtattc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val PhePro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tacgat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr AspVal Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tacgga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr GlyPhe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aatgcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn AlaTrp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acccgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr ArgCys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gttgag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val GluGlu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc agacag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg GlnAla Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc cccctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro LeuThr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgcgcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg AlaSer Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgttac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys TyrLeu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 19151920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acggag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr GluAla 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaacca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys ProGlu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtgtca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val SerVal Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acccgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr ArgAsp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag acagct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr AlaArg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atgtat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met TyrAla Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttcttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe PheSer Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gattgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp CysGln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac ctacct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu ProGln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tcactc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser LeuHis Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgcctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys LeuArg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgggcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg AlaArg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 AlaThr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 21352140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agcctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser LeuSer Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc ctactt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu LeuSer Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tgaacggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 21952200 ggccaatagg ccatcctgtt tttttccctt tttttttttc tttttttttt tttttttttt8487 tttttttttt ttttctcctt tttttttcct ctttttttcc ttttctttcc tttggtggct8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga8607 gtgctgatac tggcctctct gcagatcaag t 8638

1. A HCV self-replicating polynucleotide comprising: (a) a 5′-NTRconsisting of ACCAGC (SEQ ID NO. 8); (b) a HCV polyprotein region codingfor a HCV polyprotein; and (c) a 3′-NTR region.
 2. The HCVpolynucleotide according to claim 1, wherein said polyprotein comprisesone or more amino acid substitution selected from the group consistingof: R(1135)K, S(1148)G, S(1560)G, K(1691)R, L(1701)F, I(1984)V,T(1993)A, G(2042)C, G(2042)R, S(2404)P, L(2155)P, P(2166)L and M(2992)T.3. The HCV polynucleotide encoding a polyprotein comprising one or moreof the amino acid substitutions as defined in claim 2, and furthercomprising the amino acid substitution E(1202)G.
 4. The HCVpolynucleotide according to claim 2, wherein said substitution is aG2042C or a G2042R mutation.
 5. The HCV polynucleotide according toclaim 2, wherein said substitution is selected from the group consistingof: K(1691)R, and G(2042)C.
 6. The HCV polynucleotide according to claim2, wherein said substitution is selected from the group consisting of:R(1135)K, S(1560)G, K(1691)R, T(1993)A, G(2042)C, and P(2166)L.
 7. TheHCV polynucleotide according to claim 2, wherein said substitution isselected from the group consisting of: R(1135)K, S(1560)G, K(1691)R,T(1993)A, G(2042)C, L(2155)P, and P(2166)L.
 8. The HCV polynucleotideaccording to claim 2, wherein said substitution is selected from thegroup consisting of: E(1202)G, I(1984)V, G(2042)C, and M(2992)T.
 9. TheHCV polynucleotide according to claim 2, wherein said substitution isselected from the group consisting of: S(1148)G, E(1202)G, L(1701)F,G(2042)R, and S(2404)P.
 10. A HCV polynucleotide molecule comprising:(a) a 5′-NTR region; (b) a HCV polyprotein region coding for an HCVpolyprotein comprising one or more amino acid substitutions selectedfrom the group consisting of: R(1135)K, S(1148)G, S(1560)G, K(1691)R,L(1701)F, l(1984)V, T(1993)A, G(2042)C, G(2042)R, S(2404)P, L(2155)P,P(2166)L, and M(2992)T, and (c) a 3′-NTR region.
 11. The HCVself-replicating polynucleotide encoding a polyprotein comprising anyone of the amino acid substitutions according to claim 10, furthercomprising the amino acid substitution E(1202)G.
 12. The polynucleotideaccording to claim 10, wherein said substitution is a G2042C or a G2042Rmutation.
 13. The HCV polynucleotide according to claim 10, wherein saidsubstitution is selected from the group consisting of: K(1691)R, andG(2042)C.
 14. The HCV polynucleotide according to claim 10, wherein saidsubstitution is selected from the group consisting of: R(1135)K,S(1560)G, K(1691)R, T(1993)A, G(2042)C, and P(2166)L.
 15. The HCVpolynucleotide according to claim 10, wherein said substitution isselected from the group consisting of: ,R(1135)K, S(1560)G, K(1691)R,T(1993)A, G(2042)C, L(2155)P, and P(2166)L.
 16. The HCV polynucleotideaccording to claim 10, wherein said substitution is selected from thegroup consisting of: E(1202)G, 1(1984)V, G(2042)C, and M(2992)T.
 17. TheHCV polynucleotide according to claim 10, wherein said substitution isselected from the group consisting of: S(1148)G, E(1202)G, L(1701)F,G(2042)R, and S(2404)P.